Abstract:
Compositions and methods for the detection and treatment of cancer are provided. It has been discovered that uterine cancer cells selectively induce the survivin promoter to express a gene product. One embodiment provides a method for detecting uterine malignancy by systemic administration of a cancer-specific vector that utilizes a Survivin promoter to drive expression of a reporter gene. The expression of the reporter gene is detectable exclusively in malignant cells, for example using conventional imagining techniques.

Description:
FIELD OF THE INVENTION 
     The invention is generally directed to molecular biology, more particularly to the detection and treatment of cancer using nucleic acid constructs. 
     REFERENCE TO THE SEQUENCE LISTING 
     The Sequence Listing submitted as a text file named “GRU_2016_004_ST25.txt,” created on Sep. 30, 2016, and having a size of 27,881 bytes is hereby incorporated by reference pursuant to 37 C.F.R. §1.52(e)(5). 
     BACKGROUND OF THE INVENTION 
     Benign uterine leiomyomas (fibroids) are the most common pelvic tumor in women (estimated lifetime risk of 70 percent in white women and 80 percent in black women) (Buttram, V. C., Jr. and R. C. Reiter, Fertil Steril, 36(4): 433-445 (1981); Serden, S. P. and P. G. Brooks, J Reprod Med, 36(10): 697-699 (1991); Baird, D. et al., Am J Obstet Gynecol, 188(1):100-107 (2003)). Uterine sarcoma is rare (3 to 7 per 100,000 in the United States population) with a poor prognosis (Brooks, S. E., et al., Gynecol Oncol 93(1): 204-208 (2004)). It is well recognized that cancer is an enormous global health problem. The American Cancer Society estimates that in 2008 alone there were an estimated 12.7 million new diagnoses of cancer and 7.6 million deaths caused by cancer Reynolds, P. N., et al., Nat Biotech, 19(9): 838-842 (2001)). The time at which a cancer is detected, both at initial cancer diagnosis and during tumor recurrence, is one of the most important prognostic factors that substantially affect patient outcome, because if cancer is detected early, current treatments are likely to be more effective (Etzioni, R., et al., Nat Rev Cancer, 3(4): 243-252 (2003)). 
     Unfortunately, the majority of cancers are detected relatively late, leading to high mortality rates. These rates are expected to double by 2030 unless more effective detection strategies and treatments are developed. To stem the tremendous loss of life caused by this terrible disease, a broadly applicable tool capable of detecting cancers in their earliest stages is urgently needed. Proper differentiation between benign and malignant uterine lesions can dramatically improve the efficacy of patient treatment modalities. However, despite marvelous progress in cancer specific blood-based biomarkers, many of such biomarkers have failed clinically because of presence of limitations such as highly variable background expression from nonmalignant tissues and tumor heterogeneity. 
     Therefore, it is an object of the invention to provide compositions and methods for the early detection of cancer, in particular uterine cancer. 
     It is another object of the invention to provide compositions and methods for the treatment of cancer, in particular uterine cancer. 
     SUMMARY OF THE INVENTION 
     Compositions and methods for the detection and treatment of cancer are provided. It has been discovered that uterine cancer cells selectively induce the survivin promoter to express a gene product. One embodiment provides a method for detecting uterine malignancy by systemic administration of a cancer-specific vector that utilizes a survivin promoter to drive expression of a reporter gene. The expression of the reporter gene is detectable exclusively in malignant cells, for example using conventional imagining techniques. 
     Methods for treating uterine cancer are also provided. One method for treating uterine cancer includes administering to a subject suspected of having uterine cancer an expression vector encoding a cytotoxic agent, wherein expression of the cytotoxic agent is under the control of a survivin promoter. The vector can be administered systemically or directly into the uterus. 
     Another embodiment provides a uterine cell containing a viral vector, wherein the viral vector contains a survivin promoter. Preferred vectors are adenoviral vectors. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIGS. 1A-1C  are micrographs showing X-Gal staining of human SK-UT 1 cells after transfection with Ad-Lac Z reporter gene at multiplicity of infection 1 MOI ( FIG. 1A ), 3 MOI ( FIG. 1B ) and 5 MOI ( FIG. 1C ). 
         FIG. 2  shows the results of the luciferase assay in Leiomyosarcoma tissue, primary fibroid tissue, and myometrium transfected with constructs having a Survivin promoter at 1 MOI, Survivin promoter at 5 MOI, Secretory Leukoprotease Inhibitor promoter at 1 MOI, Secretory Leukoprotease Inhibitor promoter at 5 MOI, Heparanase promoter at 1 MOI, Heparanase promoter at 5 MOI. 
         FIG. 3  is an image of three rows of cell culture plates. The first row from top to bottom is Leiomyosarcoma, the second row is primary fibroid, and the third row is myometrium all transfected with a vector having the survivin promoter controlling expression of luciferase. MOI for each column of plates from left to right is 2, 3, 5, and 10. 
         FIG. 4  is a line graph of total photon emission/s versus MOI for LMS (♦), 1ryF (▪) or 1ryMyoF (triangle) cell lines infected with Ad5-Survivin-luc. 
         FIGS. 5A and 5B  are photographs of mice injected with 5×10 6  cells transfected with Ad5-Survivin-luc. Signal shows only in the transfected LMS lesions and near zero in the non-transfected as well as the begin leiomyoma case. (P&lt;0.0001) 
         FIG. 6A  is a photograph of mice injected with 20×10 6  transfected t-1ryF cells, transfected t-LMS cells, or LMS cells. Signal shows only on transfected t-LMS cells.  FIG. 6B  is bar graph of total photon emission (TPE) per second of LMS cells, t-1ryF cells, and t-LMS cells.  FIG. 6C  is a photograph of mice injected with LMS cells, t-LMS cells, t-LMS cells, t-1ryF cells, or t-1ryF cells 
         FIGS. 7A-7B  show images of mice with 20×10 6  cells injected intracervically. The cells are t-LMS, t-1ryF, or LMS cells.  FIG. 7A  is shows images thirty minutes post injection.  FIG. 7B  shows images one hour post injection, and  FIG. 7C  shows images post cellular implantation with the same number of cells.  FIG. 7D  is line graph of total photon emission per second versus time for from top to bottom t-LMS, t-1ryF, and LMS cells. 
         FIG. 8  is an image of mice injected with 5×10 6  cells intracervically. The cells are t-LMS, t-LMS, LMS, t-1ryF, and t-1ryF cells from left to right. Signal shows only in the infected LMS lesions and is near zero in the non-transfected as well as benign leiomyoma case. 
         FIG. 9  is a DNA sequence analysis of the 5′ flanking region of the human survivin gene (SEQ ID NO:1), including the ATG start codon and sequence up and downstream thereof (adapted from Li and Altieri, et al.,  Biochem. J,  344:305-311 (1999)). The amino acid sequence for the first exon of survivin is also shown (SEQ ID NO:3). Numbering is from the initiating ATG. Canonical Sp1, Sp1-like sites and CDE/CHR elements are boxed. Arrows indicate the position of the two main transcription start sites identified by primer extension and S1 nuclease protection experiments. An upward arrow indicates the first intron-exon boundary 
         FIG. 10  is a diagram (adapted from Zhu, et al.,  Cancer Gene Therapy,  11:256-262 (2004)) of an exemplary adenoviral vector including a survivin promoter driving expression of a lunciferase reporter open reading frame and a polyA tail cloned into the deleted E1 of an adenoviral vector backbone (pAdEasy-1). In this example, E3 is also deleted from the adenoviral vector backbone. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     I. Definitions 
     The use of the terms “a,” “an,” “the,” and similar referents in the context of describing the presently claimed invention (especially in the context of the claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. 
     Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. 
     Use of the term “about” is intended to describe values either above or below the stated value in a range of approx. +/−10%; in other embodiments the values may range in value either above or below the stated value in a range of approx. +/−5%; in other embodiments the values may range in value either above or below the stated value in a range of approx. +/−2%; in other embodiments the values may range in value either above or below the stated value in a range of approx. +/−1%. The preceding ranges are intended to be made clear by context, and no further limitation is implied. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention. 
     The term “adenovirus” as used herein refers to a non-enveloped icosahedral double-stranded DNA virus having about a linear genome of about 36 kb. 
     The term “tumor-specific promoter” as used herein refers to a promoter which is activated specifically in a tumor cell compared to a normal cell to facilitate a transcription of a gene operably linked to the promoter. 
     The term “nucleic acid construct” or “nucleic acid cassette” as used herein refers to a nucleotide sequence constructed for insertion to an expression vector. 
     The term “vector” as used herein refers to a vehicle for gene transfer as that term is understood by those skilled in the art, and includes viruses, plasmids, and the like. 
     The term “operably linked” used herein refers to the arrangement of various nucleic acid molecule elements relative to each other such that the elements are functionally connected and are able to interact with each other. 
     The term “promoter” refers to a regulatory nucleic acid sequence, typically located upstream (5′) of a gene or protein coding sequence that, in conjunction with various elements, is responsible for regulating the expression of the gene or protein coding sequence. 
     The term “expression control sequence” refers to a DNA sequence that controls and regulates the transcription and/or translation of another DNA sequence. Control sequences that are suitable for prokaryotes, for example, include a promoter, optionally an operator sequence, a ribosome binding site, and the like. Eukaryotic cells can utilize promoters, polyadenylation signals, and enhancers. 
     II. Nucleic Acid Constructs 
     Nucleic acid constructs for detecting or treating uterine cancer include, but are not limited to expression vectors under the control of a Survivin promoter. For detecting cancer cells, the constructs include a reporter gene. Constructs for treating cancer encode a cytotoxic agent or oncolytic virus. Preferred constructs are viral vectors. The virus can be selected from the group consisting of adenovirus, adeno-associated virus, retrovirus, lentivirus, herpes simplex virus, and reovirus. Preferably, the virus is an adenovirus, and most preferably an adenovirus derived from primates. Adenoviruses infect both non-dividing and dividing cells unlike retroviruses and replicate as episomal elements in the nucleus without integrating with host genome, thereby not disrupting host genome. Adenoviruses are also useful for gene therapy due to high efficacy, long and safe storage, and low restriction in inserting an exogenous gene. 
     A. Constructs for Detecting Uterine Cancer 
     Constructs for detecting uterine cancer include but are not limited to the constructs reported in Zhu, Z., et al., Cancer Gene Therapy, 11:256-262 (2004) and Houdt, W., et al., J Neurosurg, 104:583-592 (2006) both of which are incorporated by reference in their entireties. 
     A preferred nucleic acid construct is a viral vector containing a reporter gene under the control of a survivin promoter. The reporter gene can encode an enzyme or a luminescent or fluorescent gene product. An exemplary fluorescent gene product is Green Fluorescent Protein. The reporter gene can encode luciferase which catalyzes a reaction with luciferin to produce light. 
     1. Preferred Survivin Promoter Sequences 
     A nucleic acid sequence for the human Survivin gene is 
                                       (SEQ ID NO: 4)                    1   TCTAGACATG CGGATATATT CAAGCTGGGC ACAGCACAGC AGCCCCACCC CAGGCAGCTT                      61   GAAATCAGAG CTGGGGTCCA AAGGGACCAC ACCCCGAGGG ACTGTGTGGG GGTCGGGGCA                 121   CACAGGCCAC TGCTTCCCCC CGTCTTTCTC AGCCATTCCT GAAGTCAGCC TCACTCTGCT                 181   TCTCAGGGAT TTCAAATGTG CAGAGACTCT GGCACTTTTG TAGAAGCCCC TTCTGGTCCT                 241   AACTTACACC TGGATGCTGT GGGGCTGCAG CTGCTGCTCG GGCTCGGGAG GATGCTGGGG                 301   GCCCGGTGCC CATGAGCTTT TGAAGCTCCT GGAACTCGGT TTTGAGGGTG TTCAGGTCCA                 361   GGTGGACACC TGGGCTGTCC TTGTCCATGC ATTTGATGAC ATTGTGTGCA GAAGTGAAAA                 421   GGAGTTAGGC CGGGCATGCT GGCTTATGCC TGTAATCCCA GCACTTTGGG AGGCTGAGGC                 481   GGGTGGATCA CGAGGTCAGG AGTTCAATAC CAGCCTGGCC AAGATGGTGA AACCCCGTCT                 541   CTACTAAAAA TACAAAAAAA TTAGCCGGGC ATGGTGGCGG GCGCATGTAA TCCCAGCTAC                 601   TGGGGGGGCT GAGGCAGAGA ATTGCTGGAA CCCAGGAGAT GGAGGTTGCA GTGAGCCAAG                 661   ATTGTGCCAC TGCACTGCAC TCCAGCCTGG CGACAGAGCA AGACTCTGTC TCAAAAAAAA                 721   AAAAAAAAAG TGAAAAGGAG TTGTTCCTTT CCTCCCTCCT GAGGGCAGGC AACTGCTGCG                 781   GTTGCCAGTG GAGGTGGTGC GTCCTTGGTC TGTGCCTGGG GGCCACCCCA GCAGAGGCCA                 841   TGGTGGTGCC AGGGCCCGGT TAGCGAGCCA ATCAGCAGGA CCCAGGGGCG ACCTGCCAAA                 901   GTCAACTGGA TTTGATAACT GCAGCGAAGT TAAGTTTCCT GATTTTGATG ATTGTGTTGT                 961   GGTTGTGTAA GAGAATGAAG TATTTCGGGG TAGTATGGTA ATGCCTTCAA CTTACAAACG                1021   GTTCAGGTAA ACCACCCATA TACATACATA TACATGCATG TGATATATAC ACATACAGGG                1081   ATGTGTGTGT GTTCACATAT ATGAGGGGAG AGAGACTAGG GGAGAGAAAG TAGGTTGGGG                1141   AGAGGGAGAG AGAAAGGAAA ACAGGAGACA GAGAGAGAGC GGGGAGTAGA GAGAGGGAAG                1201   GGGTAAGAGA GGGAGAGGAG GAGAGAAAGG GAGGAAGAAG CAGAGAGTGA ATGTTAAAGG                1261   AAACAGGCAA AACATAAACA GAAAATCTGG GTGAAGGGTA TATGAGTATT CTTTGTACTA                1321   TTCTTGCAAT TATCTTTTAT TTAAATTGAC ATCGGGCCGG GCGCAGTGGC TCACATCTGT                1381   AATCCCAGCA CTTTGGGAGG CCGAGGCAGG CAGATCACTT GAGGTCAGGA GTTTGAGACC                1441   AGCCTGGCAA ACATGGTGAA ACCCCATCTC TACTAAAAAT ACAAAAATTA GCCTGGTGTG                1501   GTGGTGCATG CCTTTAATCT CAGCTACTCG GGAGGCTGAG GCAGGAGAAT CGCTTGAACC                1561   CGTGGCGGGG AGGAGGTTGC AGTGAGCTGA GATCATGCCA CTGCACTCCA GCCTGGGCGA                1621   TAGAGCGAGA CTCAGTTTCA AATAAATAAA TAAACATCAA AATAAAAAGT TACTGTATTA                1681   AAGAATGGGG GCGGGGTGGG AGGGGTGGGG AGAGGTTGCA AAAATAAATA AATAAATAAA                1741   TAAACCCCAA AATGAAAAAG ACAGTGGAGG CACCAGGCCT GCGTGGGGCT GGAGGGCTAA                1801   TAAGGCCAGG CCTCTTATCT CTGGCCATAG AACCAGAGAA GTGAGTGGAT GTGATGCCCA                1861   GCTCCAGAAG TGACTCCAGA ACACCCTGTT CCAAAGCAGA GGACACACTG ATTTTTTTTT                1921   TAATAGGCTG CAGGACTTAC TGTTGGTGGG ACGCCCTGCT TTGCGAAGGG AAAGGAGGAG                1981   TTTGCCCTGA GCACAGGCCC CCACCCTCCA CTGGGCTTTC CCCAGCTCCC TTGTCTTCTT                2041   ATCACGGTAG TGGCCCAGTC CCTGGCCCCT GACTCCAGAA GGTGGCCCTC CTGGAAACCC                2101   AGGTCGTGCA GTCAACGATG TACTCGCCGG GACAGCGATG TCTGCTGCAC TCCATCCCTC                2161   CCCTGTTCAT TTGTCCTTCA TGCCCGTCTG GAGTAGATGC TTTTTGCAGA GGTGGCACCC                2221   TGTAAAGCTC TCCTGTCTGA CTTTTTTTTT TTTTTTAGAC TGAGTTTTGC TCTTGTTGCC                2281   TAGGCTGGAG TGCAATGGCA CAATCTCAGC TCACTGCACC CTCTGCCTCC CGGGTTCAAG                2341   CGATTCTCCT GCCTCAGCCT CCCGAGTAGT TGGGATTACA GGCATGCACC ACCACGCCCA                2401   GCTAATTTTT GTATTTTTAG TAGAGACAAG GTTTCACCGT GATGGCCAGG CTGGTCTTGA                2461   ACTCCAGGAC TCAAGTGATG CTCCTGCCTA GGCCTCTCAA AGTGTTGGGA TTACAGGCGT                2521   GAGCCACTGC ACCCGGCCTG CACGCGTTCT TTGAAAGCAG TCGAGGGGGC GCTAGGTGTG                2581   GGCAGGGACG AGCTGGCGCG GCGTCGCTGG GTGCACCGCG ACCACGGGCA GAGCCACGCG                2641   GCGGGAGGAC TACAACTCCC GGCACACCCC GCGCCGCCCC GCCTCTACTC CCAGAAGGCC                2701   GCGGGGGGTG GACCGCCTAA GAGGGCGTGC GCTCCCGACA TGCCCCGCGG CGCGCCATTA                2761   ACCGCCAGAT TTGAATCGCG GGACCCGTTG GCAGAGGTGG CGGCGGCGGC    ATG   GGTGCCC                2821   CGACGTTGCC CCCTGCCTGG CAGCCCTTTC TCAAGGACCA CCGCATCTCT ACATTCAAGA                2881   ACTGGCCCTT CTTGGAGGGC TGCGCCTGCA CCCCGGAGCG GGTGAGACTG CCCGGCCTCC                2941   TGGGGTCCCC CACGCCCGCC TTGCCCTGTC CCTAGCGAGG CCACTGTGAC TGGGCCTCGG                3001   GGGTACAAGC CGCCCTCCCC TCCCCGTCCT GTCCCCAGCG AGGCCACTGT GGCTGGGCCC                3061   CTTGGGTCCA GGCCGGCCTC CCCTCCCTGC TTTGTCCCCA TCGAGGCCTT TGTGGCTGGG                3121   CCTCGGGGTT CCGGGCTGCC ACGTCCACTC ACGAGCTGTG CTGTCCCTTG CAGATGGCCG                3181   AGGCTGGCTT CATCCACTGC CCCACTGAGA ACGAGCCAGA CTTGGCCCAG TGTTTCTTCT                3241   GCTTCAAGGA GCTGGAAGGC TGGGAGCCAG ATGACGACCC CATGTAAGTC TTCTCTGGCC                3301   AGCCTCGATG GGCTTTGTTT TGAACTGAGT TGTCAAAAGA TTTGAGTTGC AAAGACACTT                3361   AGTATGGGAG GGTTGCTTTC CACCCTCATT GCTTCTTAAA CAGCTGTTGT GAACGGATAC                3421   CTCTCTATAT GCTGGTGCCT TGGTGATGCT TACAACCTAA TTAAATCTCA TTTGACCAAA                3481   ATGCCTTGGG GTGGACGTAA GATGCCTGAT GCCTTTCATG TTCAACAGAA TACATCAGCA                3541   GACCCTGTTG TTGTGAACTC CCAGGAATGT CCAAGTGCTT TTTTTGAGAT TTTTTAAAAA                3601   ACAGTTTAAT TGAAATATAA CCTACACAGC ACAAAAATTA CCCTTTGAAA GTGTGCACTT                3661   CACACTTTCG GAGGCTGAGG CGGGCGGATC ACCTGAGGTC AGGAGTTCAA GACCTGCCTG                3721   GCCAACTTGG CGAAACCCCG TCTCTACTAA AAATACAAAA ATTAGCCGGG CATGGTAGCG                3781   CACGCCCGTA ATCCCAGCTA CTCGGGAGGC TAAGGCAGGA GAATCGCTTG AACCTGGGAG                3841   GCGGAGGTTG CAGTGAGCCG AGATTGTGCC AATGCACTCC AGCCTCGGCG ACAGAGCGAG                3901   ACTCCGTCAT AAAAATAAAA AATTGAAAAA AAAAAAAGAA AGAAAGCATA TACTTCAGTG                3961   TTGTTCTGGA TTTTTTTCTT CAAGATGCCT AGTTAATGAC AATGAAATTC TGTACTCGGA                4021   TGGTATCTGT CTTTCCACAC TGTAATGCCA TATTCTTTTC TCACCTTTTT TTCTGTCGGA                4081   TTCAGTTGCT TCCACAGCTT TAATTTTTTT CCCCTGGAGA ATCACCCCAG TTGTTTTTCT                4141   TTTTGGCCAG AAGAGAGTAG CTGTTTTTTT TCTTAGTATG TTTGCTATGG TGGTTATACT                4201   GCATCCCCGT AATCACTGGG AAAAGATCAG TGGTATTCTT CTTGAAAATG AATAAGTGTT                4261   ATGATATTTT CAGATTAGAG TTACAACTGG CTGTCTTTTT GGACTTTGTG TGGCCATGTT                4321   TTCATTGTAA TGCAGTTCTG GTAACGGTGA TAGTCAGTTA TACAGGGAGA CTCCCCTAGC                4381   AGAAAATGAG AGTGTGAGCT AGGGGGTCCC TTGGGGAACC CGGGGCAATA ATGCCCTTCT                4441   CTGCCCTTAA TCCTTACAGT GGGCCGGGCA CGGTGGCTTA CGCCTGTAAT ACCAGCACTT                4501   TGGGAGGCCG AGGCGGGCGG ATCACGAGGT CAGGAGATCG AGACCATCTT GGCTAATACG                4561   GTGAAACCCC GTCTCCACTA AAAATACAAA AAATTAGCCG GGCGTGGTGG TGGGCGCCTG                4621   TAGTCCCAGC TACTCGGGAG GCTGAGGCAG GAGAATGGCG TGAACCCAGG AGGCGGAGCT                4681   TGCAGTGAGC CGAGATTGCA CCACTGCACT CCAGCCTGGG CGACAGAATG AGACTCCGTC                4741   TCAAAAAAAA AAAAAAAAGA AAAAAATCTT TACAGTGGAT TACATAACAA TTCCAGTGAA                4801   ATGAAATTAC TTCAAACAGT TCCTTGAGAA TGTTGGAGGG ATTTGACATG TAATTCCTTT                4861   GGACATATAC CATGTAACAC TTTTCCAACT AATTGCTAAG GAAGTCCAGA TAAAATAGAT                4921   ACATTAGCCA CACAGATGTG GGGGGAGATG TCCACAGGGA GAGAGAAGGT GCTAAGAGGT                4981   GCCATATGGG AATGTGGCTT GGGCAAAGCA CTGATGCCAT CAACTTCAGA CTTGACGTCT                5041   TACTCCTGAG GCAGAGCAGG GTGTGCCTGT GGAGGGCGTG GGGAGGTGGC CCGTGGGGAG                5101   TGGACTGCCG CTTTAATCCC TTCAGCTGCC TTTCCGCTGT TGTTTTGATT TTTCTAGAGA                5161   GGAACATAAA AAGCATTCGT CCGGTTGCGC TTTCCTTTCT GTCAAGAAGC AGTTTGAAGA                5221   ATTAACCCTT GGTGAATTTT TGAAACTGGA CAGAGAAAGA GCCAAGAACA AAATTGTATG                5281   TATTGGGAAT AAGAACTGCT CAAACCCTGT TCAATGTCTT TAGCACTAAA CTACCTAGTC                5341   CCTCAAAGGG ACTCTGTGTT TTCCTCAGGA AGCATTTTTT TTTTTTTTCT GAGATAGAGT                5401   TTCACTCTTG TTGCCCAGGC TGGAGTGCAA TGGTGCAATC TTGGCTCACT GCAACCTCTG                5461   CCTCTCGGGT TCAAGTGATT CTCCTGCCTC AGCCTCCCAA GTAACTGGGA TTACAGGGAA                5521   GTGCCACCAC ACCCAGCTAA TTTTTGTATT TTTAGTAGAG ATGGGGTTTC ACCACATTGC                5581   CCAGGCTGGT CTTGAACTCC TGACCTCGTG ATTCGCCCAC CTTGGCCTCC CAAAGTGCTG                5641   GGATTACAGG CGTGAACCAC CACGCCTGGC TTTTTTTTTT TTGTTCTGAG ACACAGTTTC                5701   ACTCTGTTAC CCAGGCTGGA GTAGGGTGGC CTGATCTCGG ATCACTGCAA CCTCCGCCTC                5761   CTGGGCTCAA GTGATTTGCC TGCTTCAGCC TCCCAAGTAG CCGAGATTAC AGGCATGTGC                5821   CACCACACCC AGGTAATTTT TGTATTTTTG GTAGAGACGA GGTTTCACCA TGTTGGCCAG                5881   GCTGGTTTTG AACTCCTGAC CTCAGGTGAT CCACCCGCCT CAGCCTCCCA AAGTGCTGAG                5941   ATTATAGGTG TGAGCCACCA CACCTGGCCT CAGGAAGTAT TTTTATTTTT AAATTTATTT                6001   ATTTATTTGA GATGGAGTCT TGCTCTGTCG CCCAGGCTAG AGTGCAGCGA CGGGATCTCG                6061   GCTCACTGCA AGCTCCGCCC CCCAGGTTCA AGCCATTCTC CTGCCTCAGC CTCCCGAGTA                6121   GCTGGGACTA CAGGCGCCCG CCACCACACC CGGCTAATTT TTTTGTATTT TTAGTAGAGA                6181   CGGGTTTTCA CCGTGTTAGC CAGGAGGGTC TTGATCTCCT GACCTCGTGA TCTGCCTGCC                6241   TCGGCCTCCC AAAGTGCTGG GATTACAGGT GTGAGCCACC ACACCCGGCT ATTTTTATTT                6301   TTTTGAGACA GGGACTCACT CTGTCACCTG GGCTGCAGTG CAGTGGTACA CCATAGCTCA                6361   CTGCAGCCTC GAACTCCTGA GCTCAAGTGA TCCTCCCACC TCATCCTCAC AAGTAATTGG                6421   GACTACAGGT GCACCCCACC ATGCCCACCT AATTTATTTA TTTATTTATT TATTTATTTT                6481   CATAGAGATG AGGGTTCCCT GTGTTGTCCA GGCTGGTCTT GAACTCCTGA GCTCACGGGA                6541   TCCTTTTGCC TGGGCCTCCC AAAGTGCTGA GATTACAGGC ATGAGCCACC GTGCCCAGCT                6601   AGGAATCATT TTTAAAGCCC CTAGGATGTC TGTGTGATTT TAAAGCTCCT GGAGTGTGGC                6661   CGGTATAAGT ATATACCGGT ATAAGTAAAT CCCACATTTT GTGTCAGTAT TTACTAGAAA                6721   CTTAGTCATT TATCTGAAGT TGAAATGTAA CTGGGCTTTA TTTATTTATT TATTTATTTA                6781   TTTATTTTTA ATTTTTTTTT TTGAGACGAG TCTCACTTTG TCACCCAGGC TGGAGTGCAG                6841   TGGCACGATC TCGGCTCACT GCAACCTCTG CCTCCCGGGG TCAAGCGATT CTCCTGCCTT                6901   AGCCTCCCGA GTAGCTGGGA CTACAGGCAC GCACCACCAT GCCTGGCTAA TTTTTGTATT                6961   TTTAGTAGAC GGGGTTTCAC CATGCTGGCC AAGCTGGTCT CAAACTCCTG ACCTTGTGAT                7021   CTGCCCGCTT TAGCCTCCCA GAGTGCTGGG ATTACAGGCA TGAGCCACCA TGCGTGGTCT                7081   TTTTAAAATT TTTTGATTTT TTTTTTTTTT GAGACAGAGC CTTGCTCTGT CGCCCAGGCT                7141   GGAGTGCAGT GGCACGATCT CAGCTCACTA CAAGCTCCGC CTCCCGGGTT CACGCCATTC                7201   TTCTGCCTCA GCCTCCTGAG TAGCTGGGAC TACAGGTGCC CACCACCACG CCTGGCTAAT                7261   TTTTTTTGGT ATTTTTATTA GAGACAAGGT TTCATCATGT TGGCCAGGCT GGTCTCAAAC                7321   TCCTGACCTC AAGTGATCTG CCTGCCTCGG CCTCCCAAAG CGCTGAGATT ACAGGTGTGA                7381   TCTACTGCGC CAGGCCTGGG CGTCATATAT TCTTATTTGC TAAGTCTGGC AGCCCCACAC                7441   AGAATAAGTA CTGGGGGATT CCATATCCTT GTAGCAAAGC CCTGGGTGGA GAGTCAGGAG                7501   ATGTTGTAGT TCTGTCTCTG CCACTTGCAG ACTTTGAGTT TAAGCCAGTC GTGCTCATGC                7561   TTTCCTTGCT AAATAGAGGT TAGACCCCCT ATCCCATGGT TTCTCAGGTT GCTTTTCAGC                7621   TTGAAAATTG TATTCCTTTG TAGAGATCAG CGTAAAATAA TTCTGTCCTT ATATGTGGCT                7681   TTATTTTAAT TTGAGACAGA GTGTCACTCA GTCGCCCAGG CTGGAGTGTG GTGGTGCGAT                7741   CTTGGCTCAC TGCGACCTCC ACCTCCCAGG TTCAAGCGAT TCTCGTGCCT CAGGCTCCCA                7801   AGTAGCTGAG ATTATAGGTG TGTGCCACCA GGCCCAGCTA ACTTTTGTAT TTTTAGTAGA                7861   GACAGGGTTT TGCCATGTTG GCTAAGCTGG TCTCGAACTC CTGGCCTCAA GTGATCTGCC                7921   CGCCTTGGCA TCCCAAAGTG CTGGGATTAC AGGTGTGAAC CACCACACCT GGCCTCAATA                7981   TAGTGGCTTT TAAGTGCTAA GGACTGAGAT TGTGTTTTGT CAGGAAGAGG CCAGTTGTGG                8041   GTGAAGCATG CTGTGAGAGA GCTTGTCACC TGGTTGAGGT TGTGGGAGCT GCAGCGTGGG                8101   AACTGGAAAG TGGGCTGGGG ATCATCTTTT TCCAGGTCAG GGGTCAGCCA GCTTTTCTGC                8161   AGCGTGCCAT AGACCATCTC TTAGCCCTCG TGGGTCAGAG TCTCTGTTGC ATATTGTCTT                8221   TTGTTGTTTT TCACAACCTT TTAGAAACAT AAAAAGCATT CTTAGCCCGT GGGCTGGACA                8281   AAAAAAGGCC ATGACGGGCT GTATGGATTT GGCCCAGCAG GCCCTTGCTT GCCAAGCCCT                8341   GTTTTAGACA AGGAGCAGCT TGTGTGCCTG GAACCATCAT GGGCACAGGG GAGGAGCAGA                8401   GTGGATGTGG AGGTGTGAGC TGGAAACCAG GTCCCAGAGC GCTGAGAAAG ACAGAGGGTT                8461   TTTGCCCTTG CAAGTAGAGC AACTGAAATC TGACACCATC CAGTTCCAGA AAGCCCTGAA                8521   GTGCTGGTGG ACGCTGCGGG GTGCTCCGCT CTAGGGTTAC AGGGATGAAG ATGCAGTCTG                8581   GTAGGGGGAG TCCACTCACC TGTTGGAAGA TGTGATTAAG AAAAGTAGAC TTTCAGGGCC                8641   GGGCATGGTG GCTCACGCCT GTAATCCCAG CACTTTGGGA GGCCGAGGCG GGTGGATCAC                8701   GAGGTCAGGA GATCGAGACC ATCCTGGCTA ACATGGTGAA ACCCCGTCTT TACTAAAAAT                8761   ACAAAAAATT AGCTGGGCGT GGTGGCGGGC GCCTGTAGTC CCAGCTACTC GGGAGGCTGA                8821   GGCAGGAGAA TGGCGTGAAC CTGGGAGGTG GAGCTTGCTG TGAGCCGAGA TCGCGCCACT                8881   GCACTCCAGC CTGGGCGACA GAGCGAGACT CCGTCTCAAA AAAAAAAAAA AAAGTAGGCT                8941   TTCATGATGT GTGAGCTGAA GGCGCAGTAG GCAGAAGTAG AGGCCTCAGT CCCTGCAGGA                9001   GACCCCTCGG TCTCTATCTC CTGATAGTCA GACCCAGCCA CACTGGAAAG AGGGGAGACA                9061   TTACAGCCTG CGAGAAAAGT AGGGAGATTT AAAAACTGCT TGGCTTTTAT TTTGAACTGT                9121   TTTTTTTGTT TGTTTGTTTT CCCCAATTCA GAATACAGAA TACTTTTATG GATTTGTTTT                9181   TATTACTTTA ATTTTGAAAC AATATAATCT TTTTTTTGTT GTTTTTTTGA GACAGGGTCT                9241   TACTCTGTCA CCCAGGCTGA GTGCAGTGGT GTGATCTTGG CTCACCTCAG CCTCGACCCC                9301   CTGGGCTCAA ATGATTCTCC CACCTCAGCT TCCCAAGTAG CTGGGACCAC AGGTGCGTGT                9361   GTTGCGCTAT ACAAATCCTG AAGACAAGGA TGCTGTTGCT GGTGATGCTG GGGATTCCCA                9421   AGATCCCAGA TTTGATGGCA GGATGCCCCT GTCTGCTGCC TTGCCAGGGT GCCAGGAGGG                9481   CGCTGCTGTG GAAGCTGAGG CCCGGCCATC CAGGGCGATG CATTGGGCGC TGATTCTTGT                9541   TCCTGCTGCT GCCTCGGTGC TTAGCTTTTG AAACAATGAA ATAAATTAGA ACCAGTGTGA                9601   AAATCGATCA GGGAATAAAT TTAATGTGGA AATAAACTGA ACAACTTAGT TCTTCATAAG                9661   AGTTTACTTG GTAAATACTT GTGATGAGGA CAAAACGAAG CACTAGAAGG AGAGGCGAGT                9721   TGTAGACCTG GGTGGCAGGA GTGTTTTGTT TGTTTTCTTT GGCAGGGTCT TGCTCTGTTG                9781   CTCAGGCTGG AGTACAGTGG CACAATCACA GCTCACTATA GCCTCGACCT CCTGGACTCA                9841   AGCAATCCTC CTGCCTCAGC CTCCCAGTAG CTGGGACTAC AGGCGCATGC CACCATGCCT                9901   GGCTAATTTT AAATTTTTTT TTTTCTCTTT TTTGAGATGG AATCTCACTC TGTCGCCCAG                9961   GCTGGAGTGC AGTGGCGTGA TCTCGGCTGA CGGCAAGCTC CGCCTCCCAG GTTCACTCCA               10021   TTCGCCTGCC TCAGCCTCCC AAGTAGCTGG GACTACAGGC GCTGGGATTA CAAACCCAAA               10081   CCCAAAGTGC TGGGATTACA GGCGTGAGCC ACTGCACCCG GCCTGTTTTG TCTTTCAATA               10141   GCAAGAGTTG TGTTTGCTTC GCCCCTACCT TTAGTGGAAA AATGTATAAA ATGGAGATAT               10201   TGACCTCCAC ATTGGGGTGG TTAAATTATA GCATGTATGC AAAGGAGCTT CGCTAATTTA               10261   AGGCTTTTTT GAAAGAGAAG AAACTGAATA ATCCATGTGT GTATATATAT TTTAAAAGCC               10321   ATGGTCATCT TTCCATATCA GTAAAGCTGA GGCTCCCTGG GACTGCAGAG TTGTCCATCA               10381   CAGTCCATTA TAAGTGCGCT GCTGGGCCAG GTGCAGTGGC TTGTGCCTGA ATCCCAGCAC               10441   TTTGGGAGGC CAAGGCAGGA GGATTCATTG AGCCCAGGAG TTTTGAGGCG AGCCTGGGCA               10501   ATGTGGCCAG ACCTCATCTC TTCAAAAAAT ACACAAAAAA TTAGCCAGGC ATGGTGGCAC               10561   GTGCCTGTAG TCTCAGCTAC TCAGGAGGCT GAGGTGGGAG GATCACTTTG AGCCTTGCAG               10621   GTCAAAGCTG CAGTAAGCCA TGATCTTGCC ACTGCATTCC AGCCTGGATG ACAGAGCGAG               10681   ACCCTGTCTC TAAAAAAAAA AAAAACCAAA CGGTGCACTG TTTTCTTTTT TCTTATCAAT               10741   TTATTATTTT TAAATTAAAT TTTCTTTTAA TAATTTATAA ATTATAAATT TATATTAAAA               10801   AATGACAAAT TTTTATTACT TATACATGAG GTAAAACTTA GGATATATAA AGTACATATT               10861   GAAAAGTAAT TTTTTGGCTG GCACAGTGGC TCACACCTGT AATCCCAGCA CTTTGGGAGG               10921   CCGTGGCGGG CAGATCACAT GAGATCATGA GTTCGAGACC AACCTGACCA ACATGGAGAG               10981   ACCCCATCTC TACTAAAAAT ACAAAATTAG CCGGGGTGGT GGCGCATGCC TGTAATCCCA               11041   GCTACTCGGG AGGCTGAGGC AGGAGAATCT CTTGAACCCG GGAGGCAGAG GTTGCGGTGA               11101   GCCAAGATCG TGCCTTTGCA CACCAGCCTA GGCAACAAGA GCGAAAGTCC GTCTCAAAAA               11161   AAAAGTAATT TTTTTTAAGT TAACCTCTGT CAGCAAACAA ATTTAACCCA ATAAAGGTCT               11221   TTGTTTTTTA ATGTAGTAGA GGAGTTAGGG TTTATAAAAA ATATGGTAGG GAAGGGGGTC               11281   CCTGGATTTG CTAATGTGAT TGTCATTTGC CCCTTAGGAG AGAGCTCTGT TAGCAGAATG               11341   AAAAAATTGG AAGCCAGATT CAGGGAGGGA CTGGAAGCAA AAGAATTTCT GTTCGAGGAA               11401   GAGCCTGATG TTTGCCAGGG TCTGTTTAAC TGGACATGAA GAGGAAGGCT CTGGACTTTC               11461   CTCCAGGAGT TTCAGGAGAA AGGTAGGGCA GTGGTTAAGA GCAGAGCTCT GCCTAGACTA               11521   GCTGGGGTGC CTAGACTAGC TGGGGTGCCC AGACTAGCTG GGGTGCCTAG ACTAGCTGGG               11581   TACTTTGAGT GGCTCCTTCA GCCTGGACCT CGGTTTCCTC ACCTGTATAG TAGAGATATG               11641   GGAGCACCCA GCGCAGGATC ACTGTGAACA TAAATCAGTT AATGGAGGAA GCAGGTAGAG               11701   TGGTGCTGGG TGCATACCAA GCACTCCGTC AGTGTTTCCT GTTATTCGAT GATTAGGAGG               11761   CAGCTTAAAC TAGAGGGAGT TGAGCTGAAT CAGGATGTTT GTCCCAGGTA GCTGGGAATC               11821   TGCCTAGCCC AGTGCCCAGT TTATTTAGGT GCTCTCTCAG TGTTCCCTGA TTGTTTTTTC               11881   CTTTGTCATC TTATCTACAG GATGTGACTG GGAAGCTCTG GTTTCAGTGT CATGTGTCTA               11941   TTCTTTATTT CCAGGCAAAG GAAACCAACA ATAAGAAGAA AGAATTTGAG GAAACTGCGA               12001   AGAAAGTGCG CCGTGCCATC GAGCAGCTGG CTGCCATGGA TTGAGGCCTC TGGCCGGAGC               12061   TGCCTGGTCC CAGAGTGGCT GCACCACTTC CAGGGTTTAT TCCCTGGTGC CACCAGCCTT               12121   CCTGTGGGCC CCTTAGCAAT GTCTTAGGAA AGGAGATCAA CATTTTCAAA TTAGATGTTT               12181   CAACTGTGCT CCTGTTTTGT CTTGAAAGTG GCACCAGAGG TGCTTCTGCC TGTGCAGCGG               12241   GTGCTGCTGG TAACAGTGGC TGCTTCTCTC TCTCTCTCTC TTTTTTGGGG GCTCATTTTT               12301   GCTGTTTTGA TTCCCGGGCT TACCAGGTGA GAAGTGAGGG AGGAAGAAGG CAGTGTCCCT               12361   TTTGCTAGAG CTGACAGCTT TGTTCGCGTG GGCAGAGCCT TCCACAGTGA ATGTGTCTGG               12421   ACCTCATGTT GTTGAGGCTG TCACAGTCCT GAGTGTGGAC TTGGCAGGTG CCTGTTGAAT               12481   CTGAGCTGCA GGTTCCTTAT CTGTCACACC TGTGCCTCCT CAGAGGACAG TTTTTTTGTT               12541   GTTGTGTTTT TTTGTTTTTT TTTTTTGGTA GATGCATGAC TTGTGTGTGA TGAGAGAATG               12601   GAGACAGAGT CCCTGGCTCC TCTACTGTTT AACAACATGG CTTTCTTATT TTGTTTGAAT               12661   TGTTAATTCA CAGAATAGCA CAAACTACAA TTAAAACTAA GCACAAAGCC ATTCTAAGTC               12721   ATTGGGGAAA CGGGGTGAAC TTCAGGTGGA TGAGGAGACA GAATAGAGTG ATAGGAAGCG               12781   TCTGGCAGAT ACTCCTTTTG CCACTGCTGT GTGATTAGAC AGGCCCAGTG AGCCGCGGGG               12841   CACATGCTGG CCGCTCCTCC CTCAGAAAAA GGCAGTGGCC TAAATCCTTT TTAAATGACT               12901   TGGCTCGATG CTGTGGGGGA CTGGCTGGGC TGCTGCAGGC CGTGTGTCTG TCAGCCCAAC               12961   CTTCACATCT GTCACGTTCT CCACACGGGG GAGAGACGCA GTCCGCCCAG GTCCCCGCTT               13021   TCTTTGGAGG CAGCAGCTCC CGCAGGGCTG AAGTCTGGCG TAAGATGATG GATTTGATTC               13081   GCCCTCCTCC CTGTCATAGA GCTGCAGGGT GGATTGTTAC AGCTTCGCTG GAAACCTCTG               13141   GAGGTCATCT CGGCTGTTCC TGAGAAATAA AAAGCCTGTC ATTTCAAACA CTGCTGTGGA               13201   CCCTACTGGG TTTTTAAAAT ATTGTCAGTT TTTCATCGTC GTCCCTAGCC TGCCAACAGC               13261   CATCTGCCCA GACAGCCGCA GTGAGGATGA GCGTCCTGGC AGAGACGCAG TTGTCTCTGG               13321   GCGCTTGCCA GAGCCACGAA CCCCAGACCT GTTTGTATCA TCCGGGCTCC TTCCGGGCAG               13381   AAACAACTGA AAATGCACTT CAGACCCACT TATTTATGCC ACATCTGAGT CGGCCTGAGA               13441   TAGACTTTTC CCTCTAAACT GGGAGAATAT CACAGTGGTT TTTGTTAGCA GAAAATGCAC               13501   TCCAGCCTCT GTACTCATCT AAGCTGCTTA TTTTTGATAT TTGTGTCAGT CTGTAAATGG               13561   ATACTTCACT TTAATAACTG TTGCTTAGTA ATTGGCTTTG TAGAGAAGCT GGAAAAAAAT               13621   GGTTTTGTCT TCAACTCCTT TGCATGCCAG GCGGTGATGT GGATCTCGGC TTCTGTGAGC               13681   CTGTGCTGTG GGCAGGGCTG AGCTGGAGCC GCCCCTCTCA GCCCGCCTGC CACGGCCTTT               13741   CCTTAAAGGC CATCCTTAAA ACCAGACCCT CATGGCTGCC AGCACCTGAA AGCTTCCTCG               13801   ACATCTGTTA ATAAAGCCGT AGGCCCTTGT CTAAGCGCAA CCGCCTAGAC TTTCTTTCAG               13861   ATACATGTCC ACATGTCCAT TTTTCAGGTT CTCTAAGTTG GAGTGGAGTC TGGGAAGGGT               13921   TGTGAATGAG GCTTCTGGGC TATGGGTGAG GTTCCAATGG CAGGTTAGAG CCCCTCGGGC               13981   CAACTGCCAT CCTGGAAAGT AGAGACAGCA GTGCCCGCTG CCCAGAAGAG ACCAGCAAGC               14041   CAAACTGGAG CCCCCATTGC AGGCTGTCGC CATGTGGAAA GAGTAACTCA CAATTGCCAA               14101   TAAAGTCTCA TGTGGTTTTA TCTACTTTTT TTTTCTTTTT CTTTTTTTTT GAGACAAGGC               14161   CTTGCCCTCC CAGGCTGGAG TGCAGTGGAA TGACCACAGC TCACCGCAAC CTCAAATTCT               14221   TGCGTTCAAG TGAACCTCCC ACTTTAGCCT CCCAAGTAGC TGGGACTACA GGCGCACGCC               14281   ATCACACCCG GCTAATTGAA AAATTTTTTT TTTTGTTTAG ATGGAATCTC ACTTTGTTGC               14341   CCAGGCTGGT CTCAAACTCC TGGGCTCAAG TGATCATCCT GCTTCAGCGT CCGACTTGTT               14401   GGTATTATAG GCGTGAGCCA CTGGGCCTGA CCTAGCTACC ATTTTTTAAT GCAGAAATGA               14461   AGACTTGTAG AAATGAAATA ACTTGTCCAG GATAGTCGAA TAAGTAACTT TTAGAGCTGG               14521   GATTTGAACC CAGGCAATCT GGCTCCAGAG CTGGGCCCTC ACTGCTGAAG GACACTGTCA               14581   GCTTGGGAGG GTGGCTATGG TCGGCTGTCT GATTCTAGGG AGTGAGGGCT GTCTTTAAAG               14641   CACCCCATTC CATTTTCAGA CAGCTTTGTC AGAAAGGCTG TCATATGGAG CTGACACCTG               14701   CCTCCCCAAG GCTTCCATAG ATCCTCTCTG TACATTGTAA CCTTTTATTT TGAAATGAAA               14761   ATTCACAGGA AGTTGTAAGG CTAGTACAGG GGATCC.            
The “ATG” start codon is underlined and bolded in the sequence above. See also GenBank Accession No. GenBank: U75285.1, Version: U75285.1 GI:2315862,  Homo sapiens apoptosis inhibitor survivin gene, complete cds , and Ambrosini, et al.,  Nature Medicine,  3:917 -921 (1997), each of which is specifically incorporated by reference herein in its entirety.
 
     As illustrated in  FIG. 9 , for reference purposes, in addition to the nucleotide numbering provided herein and in the sequence listing, nucleotides beginning with, and downstream of, the “A” in a start codon can be referred to by counting positively from +1, while sequences upstream of “A” in the start codon can be referred to by counting negatively in the reverse direction from −1 beginning with the nucleotide immediately adjacent to and upstream of the start codon “A.” Thus, in the sequence above, the underlined “ATG” can be referred to as nucleotides +1, +2, +3, respectively, while the “C” adjacent to and upstream of the “ATG,” can be referred to as nucleotide −1. 
     The disclosed constructs for use in detection and treatment of uterine cancer typically include a survivin promoter. For example, the survivin promoter can include the expression control sequence of the nucleic acid sequence of SEQ ID NO:4, or SEQ ID NO:1, illustrated in  FIG. 9 , adapted from Li and Altieri, et al.,  Biochem. J,  344:305-311 (1999), which is specifically incorporated by reference in its entirety. 
                                       (SEQ ID NO: 1)                   1   AAATTGACAT CGGGCCGGGC GCAGTGGCTC ACATCTGTAA TCCCAGCACT TTGGGAGGCC                     61   GAGGCAGGCA GATCACTTGA GGTCAGGAGT TTGAGACCAG CCTGGCAAAC ATGGTGAAAC                121   CCCATCTCTA CTAAAAATAC AAAAATTAGC CTGGTGTGGT GGTGCATGCC TTTAATCTCA                181   GCTACTCGGG AGGCTGAGGC AGGAGAATCG CTTGAACCCG TGGCGGGGAG GAGGTTGCAG                241   TGAGCTGAGA TCATGCCACT GCACTCCAGC CTGGGCGATA GAGCGAGACT CAGTTTCAAA                301   TAAATAAATA AACATCAAAA TAAAAAGTTA CTGTATTAAA GAATGGGGGC GGGGTGGGAG                361   GGGTGGGGAG AGGTTGCAAA AATAAATAAA TAAATAAATA AACCCCAAAA TGAAAAAGAC                421   AGTGGAGGCA CCAGGCCTGC GTGGGGCTGG AGGGCTAATA AGGCaAGGCC TCTTATCTCT                481   GGCCATAGAA CCAGAGAAGT GAGTGGATGT GATGCCCAGC TCCAGAAGTG ACTCCAGAAC                541   ACCCTGTTCC AAAGCAGAGG ACACACTGAT TTTTTTTTTA ATAGGCTGCA GGAUTTACTG                601   TTGGTGGGAC GCCCTGCTTT GCGAAGGGAA AGGAGGAGTT TGCCCTGAGC ACAGGCCCCC                661   ACCCTCCACT GGGCTTTCCC CAGCTCCCTT GTCTTCTTAT CACGGTAGTG GCCCAGTCCC                721   TGGCCCCTGA CTCCAGAAGG TGGCCCTCCT GGAAACCCAG GTCGTGCAGT CAACGATGTA                781   CTCGCCGGGA CAGCGATGTC TGCTGCACTC CATCCCTCCC CTGTTCATTT GTCCTTCATG                841   CCCGTCTGGA GTAGATGCTT TTTGCAGAGG TGGCACCCTG TAAAGCTCTC CTGTCTGACT                901   TTTTTTTTTT TTTTAGACTG AGTTTTGCTC TTGTTGCCTA GGCTGGAGTG CAATGGCACA                961   ATCTCAGCTC ACTGCACCCT CTGCCTCCCG GGTTCAAGCG ATTCTCCTGC CTCAGCCTCC               1021   CGAGTAGTTG GGATTACAGG CATGCACCAC CACGCCCAGC TAATTTTTGT ATTTTTAGTA               1061   GAGACAAGGT TTCACCGTGA TGGCCAGGCT GGTCTTGAAC TCCAGGACTC AAGTGATGCT               1141   CCTGCCTAGG CCTCTCAAAG TGTTGGGATT ACAGGCGTGA GCCACTGCAC CCGGCCTGCA               1201   CGCGTTCTTT GAAAGCAGTC GAGGGGGCGC TAGGTGTGGG CAGGGACGAG CTGGCGCGGC               1261   GTCGCTGGGT GCACCGCGAC CACGGGCAGA GCCACGCGGC GGGAGGACTA CAACTCCCGG               1321   CACACCCCGC GCCGCCCCGC CTCTACTCCC AGAAGGCCGC GGGGGGTGGA CCGCCTAAGA               1361   GGGCGTGCGC TCCCGACATG CCCCGCGGCG CGCCATTAAC CGCCAGATTT GAATCGCGGG               1441   ACCCGTTGGC AGAGGTGGCG GCGGCGGC   AT G   GGTGCCCCG ACGTTGCCCC CTGCCTGGCA               1501   GCCCTTTCTC AAGGACCACC GCATCTCTAC ATTCAAGAAC TGGCCCTTCT TGGAGGGCTG               1561   CGCCTGCACC CCGGAGCGGG TGAGACTGCC CGGCC.            
The “ATG” start codon is underlined and bolded in the sequence above.
 
     In some embodiments, the survivin promoter includes a 260 by DNA fragment of the human survivin promoter according to nucleotides −230 to +30 of  FIG. 9  (SEQ ID NO:1), or an expression controlling fragment thereof. Nucleotides −230 to +30 of  FIG. 9  (SEQ ID NO:1) are: 
                                       (SEQ ID NO: 5)                  1   GGCAGGGACG AGCTGGCGCG GCGTCGCTGG GTGCACCGCG ACCACGGGCA GAGCCACGCG                    61   GCGGGAGGAC TACAACTCCC GGCACACCCC GCGCCGCCCC GCCTCTACTC CCAGAAGGCC               121   GCGGGGGGTG GACCGCCTAA GAGGGCGTGC GCTCCCGACA TGCCCCGCGG CGCGCCATTA               181   ACCGCCAGAT TTGAATCGCG GGACCCGTTG GCAGAGGTGG CGGCGGCGGC    ATG   GGTGCCC               241   CGACGTTGCC CCCTGCCTGG.            
The “ATG” start codon is underlined and bolded in the sequence above. In some embodiments, the expression controlling fragment includes nucleotides −230 to −1 of  FIG. 9  (SEQ ID NO:1), which are:
 
     
       
         
               
               
             
               
               
               
             
           
               
                 (SEQ ID NO: 6) 
                   
               
             
          
           
               
                   1 
                 GGCAGGGACG AGCTGGCGCG GCGTCGCTGG GTGCACCGCG ACCACGGGCA GAGCCACGCG 
                   
               
               
                   
               
               
                  61 
                 GCGGGAGGAC TACAACTCCC GGCACACCCC GCGCCGCCCC GCCTCTACTC CCAGAAGGCC 
               
               
                   
               
               
                 121 
                 GCGGGGGGTG GACCGCCTAA GAGGGCGTGC GCTCCCGACA TGCCCCGCGG CGCGCCATTA 
               
               
                   
               
               
                 181 
                 ACCGCCAGAT TTGAATCGCG GGACCCGTTG GCAGAGGTGG CGGCGGCGGC. 
               
             
          
         
       
     
     In some embodiments, the survivin promoter is derived from a homologue or orthologue of the promoter of SEQ ID NO:1 or 4. The survivin promoter can be an expression controlling nucleic acid sequence having at least 60, 70, 80, 85, 90, 95, 96, 97, 98, or 99 percent sequence identity to the promoter of SEQ ID NO:1, or to a functional fragment thereof, such as nucleotides −230 to +30 of  FIG. 9  (SEQ ID NO:1), which is SEQ ID NO:5, or −230 to −1 of  FIG. 9  (SEQ ID NO:1), which is SEQ ID NO:6. 
     2. Preferred Reporters Genes 
     The disclosed constructs for detection of uterine cancer typically include a survivin promoter operably linked to a report. Suitable reporters are well known in the art, and include, but are not limited to, bacterial GUS gene, the firefly luciferase gene, and the cyan, green, red, and yellow fluorescent protein genes. These examples, are non-limiting, as the reporter can be any gene for which an easy and reliable assay is available can serve as the reporter gene. One of skill in the art knows which reporters are suitable or preferred for in vivo applications, ex vivo applications, or both. 
     In a particular embodiment, the reporter gene is luciferase gene of pGL3/Basic (Promega, catalog number E1751), GenBank accession no. U47295.2 (Cloning vector pGL3-Basic, complete sequence), which is specifically incorporated by reference in its entirety and provides the luciferase polypeptide sequence: 
                     (SEQ ID NO: 2)       MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVD               ITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGV               AVAPANDIYNERELLNSMGISQPTVVEVSKKGLQKILNVQKKLPIIQKII               IMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSG               STGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGEGME               TTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTL               IDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAIL               ITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG               YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVA               PAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVD               YVASQVITAKKLRGGVVEVDEVPKGLTGKLDARKIREILIKAKKGGKIAV.            
Accordingly, in some embodiments the reporter gene includes a nucleic acid sequence encoding SEQ ID NO:2, for example, the nucleic acid sequence including nucleotides 88-1740 of GenBank accession no. U47295.2.
 
     3. Vectors 
     The disclosed constructs including a survivin promoter can be inserted, using known methods, into any suitable expression vector. In particular preferred embodiments, the vector is recombinant adenoviral vector. Suitable adenoviral vectors and methods of cloning expression constructs into them are known in the art. 
     In a particular embodiment, the vector is a human Ad5 adenovirus. Sequences for Ad5 are known in the art, and can be used as the backbone for survivin driven reporter constructs disclosed herein. See, for example, Genbank accession number M73260.1 (Mastadenovirus h5 gene, complete genome), which is specifically incorporated by reference herein in its entirety. In some embodiments, E1, E3, or both are deleted or substituted (He, et al.,  Proc Natl Acad Sci USA.,  95(5):2509-14 (1998)). For example, in some embodiments, an E1 deletion is a deletion of nucleotides 455-3512, or a fragment thereof. In some embodiments, an E3 deletion is a deletion of 28587-30464, or a fragment thereof. 
     4. Method of Making Constructs and Exemplary Constructs 
     Recombinant vectors that express a reporter operably linked to a survivin expression control sequence can be constructed according to methods that are known in the art. For example, in some embodiments, the vector includes a nucleic acid sequence encoding a survivin promoter operably linked to a nucleic acid sequence encoding a firefly luciferase open reading frame. The vector can be an adenoviral vector prepared according to known materials and methods. See, for example, the AdEasy system (Agilent Technologies, Quantum, etc.) and He, et al.,  Proc Natl Acad Sci U.S.A.,  95(5):2509-14 (1998). In a specific embodiment, a construct including a luciferase reporter gene from pGL3Basic and a simian virus 40 polyadenylation (SV40 poly-A) signal driven by a survivin promoter, are cloned into the E1-deleted region of the adenoviral vector backbone using the AdEasy system. The survivin promoter is a 260 by DNA fragment of the survivin promoter (nucleotides −230 to +30 according to Li and Altieri, et al.,  Biochem. J,  344:305-311 (1999) 
     In particular embodiments, the survivin promoter is PCR amplified with or without 5′ and 3′ ends containing restriction sites suitable for cloning, or the fragment is excised by restriction digestion from a plasmid containing the promoter. For example, BamHI/Hind III can be used to recover the 260 by sequence from pLuc-cycl.2 (Li and Altieri, et al.,  Biochem. J,  344:305-311 (1999), which is specifically incorporated by reference herein in its entirety). The promotor can optionally be subcloned into an intermediate vector, such as PBS IISK(+) vector (Stratagene, La Jolla, Calif.), for example to utilize alternative restriction sites, or to add or subtract other expression construct elements if-needed. For example, following subcloning of BamHI/Hind III fragment of pLuc-cycl.2, a SacI/HindIII fragment can be cloned into pGL3/Basic (Promega, catalog number E1751) to a construct with a survivin promoter operably linked to a luciferase open reading frame. 
     It will be appreciated that the foregoing is an exemplary preparation, and steps can be omitted, substituted, or added as is known in the art. For example, any of the subcloning steps can include PCR application of the desired fragment; alternative vectors can be utilized for an alternative luciferase sequences; an alternative reporter gene can be substituted for luciferase; the survivin promoter can include more or fewer nucleotides or can be an alternative promoter sequence compared to the 260 bp promoter sequence of pLuc-cycl.2 (e.g., having one or more polymorphisms, etc.); and/or other eukaryotic expression regulatory elements can be added or subtracted. 
     Once the expression construct is a prepared, it can be inserted into an adenoviral expression system. In some embodiments, the construct is subcloned into shuttle vector (e.g., pShuttle vector (Quantum, Montreal, Quebec, Canada)), and cloned into adenoviral expression system by homologous recombination, though alternative methods of preparing adenoviral vectors well known and can be substituted. In the specific embodiment described above, a KpnI/SalI fragment from pGL3B Survivin can be subcloned into pShuttle vector to create pShuttleGL3BSurvivin and homologous recombination can be performed in BJ5183 cells in accordance with the AdEasy System to create a recombinant adenovirus, ( FIG. 10 ), in which the luciferase reporter expression is driven by the survivin promoter. See also, Zhu, et al.,  Cancer Gene Therapy,  11:256-262 (2004), and Houdt, et al.,  J. Neurosurg.,  104:583-592 (2006). 
     Other materials for preparing the disclosed constructs are also available and include, for example, BIRC5 (NM_001168) Human cDNA ORF Clone (Origene Catalogue Number RC205935), which is a BIRC5 (Myc-DDK-tagged)-Human baculoviral IAP repeat containing 5 (BIRC5/Survivin), transcript variant 1; pBS Survivin (Addgene Plasmid #19233), which is a pBluescript vector backbone including a  G. gallus  (chicken) survivin insert according to GenBank: FG356243.1 (PC/PO 2-94 Embryonic chicken perichondrium/periosteum library  Gallus gallus  cDNA clone PC/PO 2-94 5-similar to Baculoviral IAP repeat-containing 5 (survivin) (BIRC5), transcript variant 1, mRNA sequence) and Gene ID: 374078 (BIRC5 baculoviral IAP repeat containing 5 [ Gallus gallus  (chicken)]), each of which are specifically incorporated by reference in their entireties; and Luciferase-pcDNA3 (Addgene Plasmid #18964) which is a pcDNA3 including a Firefly Luciferase insert. Nucleic acid and protein sequences of all of the foregoing references, accession numbers, and reagents (e.g., plasmids and other vectors) are specifically incorporated by reference in their entireties. 
     B. Constructs for Treating Uterine Cancer 
     Constructs for treating uterine cancer include expression vectors under the control of a survivin promoter. The vectors can be prepared using materials and methods including those described above for the report constructs and others known in the art. However, it will be appreciated the therapeutic constructs optionally include a reporter gene (e.g., a reporter is not required for therapeutic constructs), and may further include additional elements not present in reporter constructs. The expression vectors can encode tumor suppressor genes, cytotoxic genes, cytostatic genes, cytokines, suicide genes, oncolytic virus and antigen-encoding genes. Examples of tumor suppressor genes include WT1, p53, p16, Rb, BRCA1, and LK8. 
     One embodiment provides a construct that expresses an oncolytic virus under the control of a survivin promoter. Oncolytic virus (OV) therapy is based on selective replication of viruses in cancer cells and their subsequent spread within a tumor without causing damage to normal tissue. Typically, OVs fall into two classes: (i) viruses that naturally replicate preferentially in cancer cells and are nonpathogenic in humans often due to elevated sensitivity to innate antiviral signaling or dependence on oncogenic signaling pathways. These include autonomous parvoviruses, myxoma virus (MYXV; poxvirus), Newcastle disease virus (NDV; paramyxovirus), reovirus, and Seneca valley virus (SVV; picornavirus); and (ii) viruses that are genetically manipulated for use as vaccine vectors, including measles virus (MV; paramyxovirus), poliovirus (PV; picornavirus), and vaccinia virus (VV; poxvirus), and/or those genetically engineered with mutations/deletions in genes required for replication in normal but not in cancer cells including adenovirus (Ad), herpes simplex virus (HSV), VV, and vesicular stomatitis virus (VSV; rhabdovirus). 
     C. Pharmaceutical Compositions 
     The nucleic acid constructs can be combined with a pharmaceutically acceptable excipient or carrier to form a pharmaceutically acceptable composition. Examples of suitable excipients or carriers include but are not limited to water, salt water, alcohol, lipid, wax, buffer solution, solid carrier such as mannitol, lactose, starches, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, and magnesium carbonate, or biodegradable microsphere (e.g., polylactate polyglycolate). 
     The compositions may be provided in the form of single dose or multi-dose container such as sealed ampule or vial. Preferably, such container may be sealed so as to conserve aseptic condition of pharmaceutical formulations before using. In general, the formulation may be preserved as suspension, fluid, and emulsion in oil or aqueous vehicle. Further, the pharmaceutical formulation may be preserved under freeze drying conditions. 
     The pharmaceutical compositions may be administered with site-specific injection or intravenous injection. Site-specific injection includes, for example, intraperitoneal injection, intrapleural injection, intrathecal injection, intraarterial injection, intratumoral injection or local application. The preferred method is intravenous injection. 
     It should be understood that the suitable amount of the nucleic acid construct actually administered ought to be determined in light of various relevant factors including the condition to be treated, the age and weight of the individual patient, food, administration time, excretion rate, the severity of the patient&#39;s symptom and reaction susceptibility; and, therefore, the above dose should not be intended to limit the scope of the invention in any way. Generally, the adenoviral vector contained in the pharmaceutical composition may be administered in an appropriate physiologically acceptable carrier at a dose of about 10 4  to about 10 14  vp/mL. The multiplicity of infection may be generally in the range of 0.001 to 100, preferably 5, 10, 20, or 50. If administered as a polynucleotide construct, about 0.01 to 1000 μg/kg of an adenoviral vector can be administered. The adenoviral vector may be administered one or more time, depending upon the intended use and the immune response potential of the host, and may also be administered as multiple, simultaneous injections. If an immune response is undesirable, the immune response may be diminished by employing a variety of immunosuppressants, or by employing a technique such as an immunoadsorption procedure (e.g., immunoapheresis) that removes adenovirus antibody from the blood, so as to permit repetitive administration, without a strong immune response. 
     The composition may be used as the single therapy. But it may be combined with other anti-tumor protocols, such as conventional chemotherapy or radiation therapy for treating cancer. The chemotherapy drug which can be used with composition of the present invention encompasses paclitaxel, cisplatin, carboplatin, procarbazine, mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, bisulfan, nitrosourea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide, tamoxifen, taxol, transplatinum, 5-fluorouracil, vincristin, vinblastin and methotrexate. The radiation therapy which can be used with the composition of the present invention may be X-ray irradiation and .gamma.-ray irradiation, etc. 
     The adenovirus produced from the adenoviral vector shows high oncolytic effect in tumor cells, while low effect in normal cells from in vitro and in vivo experiments. Thus, the viral vector comprising the tumor-specific promoter may be used for treating a cancer. 
     III. Methods for Detecting Uterine Cancer 
     One method for detecting uterine cancer includes contacting a cell or cells suspected of being cancerous with a vector containing a reporter gene, wherein the expression of the reporter gene is under the control of a survivin promoter. In a preferred embodiment, the vector is an adenoviral vector and the reporter gene encodes luciferase. 
     Another method provides administering to a subject suspected of having uterine cancer, an effective amount of a vector containing a reporter gene, wherein expression of the reporter gene is under the control of a survivin promoter. The vector can be administered systemically or directly into the uterus. 
     One embodiment provides a method for distinguishing cancerous uterine cells from non-cancerous uterine cells by contacting a population of uterine cells containing a mixture of cancerous and non-cancerous uterine cells with a vector containing a reporter gene, wherein the expression of the reporter gene is under the control of a survivin promoter, and detecting expression of the reporter gene wherein, expression of the reporter gene indicates that the cell expressing the reporter gene is cancerous. The contacting can be in vivo or ex vivo. Accordingly, methods of detecting uterine cancer in the subject and in tissue samples isolated from a subject are both specifically disclosed. 
     IV. Methods for Treating Uterine Cancer 
     One method for treating uterine cancer includes contacting a cell or cells suspected of being cancerous with a vector containing a gene that encodes tumor suppressor genes, cytotoxic genes, cytostatic genes, cytokines, suicide genes, oncolytic virus and antigen-encoding genes. Examples of tumor suppressor genes include WT1, p53, p16, Rb, BRCA1, and LK8. The expression of the gene is under the control of a survivin promoter. In a preferred embodiment, the vector is an adenoviral vector encoding an oncolytic virus, wherein the expression of the oncolytic virus is under the control of a survivin promoter. 
     Another method provides administering to a subject suspected of having uterine cancer, an effective amount of a vector encoding a cytotoxic agent, wherein expression of the cytotoxic agent is under the control of a survivin promoter. The vector can be administered systemically or directly into the uterus. 
     EXAMPLES 
     Example 1 
     X-Gal Staining 
     Materials and Methods 
     All procedures performed on animals were approved by Georgia Regents University&#39;s Institutional Animal Care and Use Committee and were within the guidelines of humane care of laboratory animals. Materials and methods used in viral vector construction, cell culture, and in vitro transfection experiments, s.c. Tumor and intrauterine models, and local vector administration, plasma collection, and statistical analyses are detailed herein materials and methods. 
     Recombinant Adenovirus and Promoters 
     Large-scale production of adenovirus vectors was performed as we have described previously with a typical batch yield of 2×10 10  plaque-forming units (PFU)/ml (Al-Hendy, A., et al., Am J Obstet Gynecol, 182(3): 553-559 (2000)). Ad vectors used in this study are listed in Table 1. 
     
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Description of the Adenovirus vectors used in this study 
               
             
          
           
               
                   
                   
                   
                 Modification 
                   
               
               
                 No. 
                 virus 
                 Promoter 
                 Site 
                 Virus description 
               
               
                   
               
               
                 1 
                 Ad5-luc 
                 CMV 
                 WILD 
                 E1/E3 deleted, a 
               
               
                   
                   
                   
                   
                 luciferase gene under the 
               
               
                   
                   
                   
                   
                 CMV promoter in place of 
               
               
                   
                   
                   
                   
                 E1 
               
               
                 2 
                 Ad5-LacZ 
                 CMV 
                 WILD 
                 E1/E3 deleted, a luciferase 
               
               
                   
                   
                   
                   
                 gene under the CMV 
               
               
                   
                   
                   
                   
                 promoter in place of E1 
               
               
                 3 
                 Ad5-survivin-luc 
                 Survivin 
                 Promoter 
                 E1/E3 deleted, a luciferase 
               
               
                   
                   
                   
                   
                 gene under the Survivin 
               
               
                   
                   
                   
                   
                 promoter in place of E1 
               
               
                 4 
                 Ad5-heparanase- 
                 Heparanase 
                 Promoter 
                 E1/E3 deleted, a luciferase 
               
               
                   
                 luc 
                   
                   
                 gene under the heparanase 
               
               
                   
                   
                   
                   
                 promoter in place of E1 
               
               
                 5 
                 Ad5-SLPI-uc 
                 Secretory 
                 Promoter 
                 E1/E3 deleted, a luciferase 
               
               
                   
                   
                 leukoprotease 
                   
                 gene under the SLPI 
               
               
                   
                   
                 Inhibitor 
                   
                 promoter in place of E1 
               
               
                   
                   
                 (SLPI) 
               
               
                   
               
               
                 No. 1 is described in Krasnykh, V. N., et al., J Virol 70(10): 6839-6846 (1996). 
               
               
                 No. 2 is described in Franklin, R., M. Quick and G. Haase, Gene Ther 6(8): 1360-1367 (1999). 
               
               
                 No. 3 is described in Van Houdt, W. J., et al., J Neurosurg 104(4): 583-592 (2006). 
               
               
                 No. 4 is described in Breidenbach, M., et al., Cancer Lett 240(1): 114-122 (2006). 
               
               
                 No. 5 is described in Barker, S. D., et al., Gene Ther 10(14): 1198-1204 (2003).  Cell cultures   
               
             
          
         
       
     
     For experimental models, the human leiomyosarcoma cell line SKUT-1 was used and purchased from American type Culture collection (ATCC® HTB-114™) that is derived from 75 years old Caucasian female originating from grade III, mesodermal tumor (mixed); consistent with uterine leiomyosarcoma. 
     The cells are considered biosafety level 1 based on U.S. Public Health Service Guidelines. Primary cultures of human leiomyoma cells were derived from fibroid tumors removed during hysterectomies. Human leiomyoma tissues were collected according to the policies of the Institutional Review Board of Georgia Regents University, Augusta, Ga., USA, and used to establish primary fibroid (1ry F) cells, as described previously (Rauk, P. N., et al, Am J Obstet Gynecol 173(2): 571-577 (1995); Al-Hendy, A., et al., Am J Obstet Gynecol 191(5): 1621-1631 (2004)). To represent normal cells (controls), we used a human myometrial cell line (Myo N); this cell line was cultured and maintained as described previously (Carney, M. E. Hawaii Med J 61(12): 283-286 (2002)). 
     X Gal Staining of Fixed Leiomyosarcoma Cells 
     This experiment was done to evaluate the susceptibility of SKUT-1 cells to transfection by wild type Adenovirus serotype 5 with B-Galactosidase as a reporter gene. Three different multiplicities of infection (MOI) of 1, 3, and 5 were used. The viral particles were mixed with cell culture media followed by 1 hour of mild shaking then regular cell culture conditions were applied. 24 hours later, X-Gal staining was performed on the cells. 
     Results 
     Human Leiomyosarcoma cells (LMS) are susceptible to wild type adenovirus transfection by X-Gal Staining. The transgene of the bacterial enzyme β-galactosidase can be easily located with a LacZ stain using the artificial substrate X-Gal, which turns blue when it is cleaved by B-Galactosidase. Culture media was aspirated off and washed SKUT-1 cells 1× with cold PBS, fixed the cells on ice with ˜5 mL glutaraldehyde (1:100 dilution of stock in PBS) for 5 min, rinse the cells 3× with PBS for 4 min. per wash, dilute 25× stock of X-Gal into the staining solution (final 1 mg/mL of X-gal), add ˜5 mL of the X-gal staining solution to the cells and incubate at 37° C. for 24 hours. Cells were checked every 4 hours to determine whether cells were turning blue. ( FIGS. 1A-1C ) Transfection of Leiomyosarcoma cells by Ad-lacZ, X-Gal staining of human SK-UT 1 cells after transfection with Adenovirus with Ad-Lac Z reporter gene at multiplicity of infection 1 MOI ( FIG. 1A ), 3 MOI ( FIG. 1B ) and 5 MOI ( FIG. 1C ). X-gal is an analogue of lactose and therefore hydrolyzed by the β-galactosidase enzyme giving intensely blue products. 
     Example 2 
     Screening for Sarcoma Specific Gene Expression 
     Materials and Methods 
     Luciferase Assay 
     To screen the 3 promoters of interest for their sarcoma specific gene expression potential, #3 60 mm 2  cell culture dishes of SKUT-1 cells at 70% confluence were transfected with 3 different adenoviral constructs which are Ad Survivin, Ad Heparanase, and Ad SLPI all at the same MOI the same technique as described above and used luciferase assay to differentiate between gene expression levels under the control of the 3 used promoters. Growth medium was removed from cultured cells, and the cells were rinsed in 1× PBS. Then without dislodging cells, as much of the final wash was removed as possible. 400 ul volume of 1× cell lysis buffer was dispensed (CCLR) into each culture vessel then attached cells were scraped from the dish, and the cells and solution were transferred to a microcentrifuge tube. Debris was separated by brief centrifugation, and the supernatant was transferred to a new tube. 20 μl of cell lysate was used with 100 μl of luciferase assay reagent and measured the light produced by Synergy HT microplate reader utilizing Gen-5 software for bioluminescence detection. The classic luciferase assay was used to compare the degree of reporter gene expression in leiomyosarcoma cells under different promoters. 
     Results 
     Screening of reporter gene expression in LMS cells under different promoters reveals Ad-Survivin as a malignancy specific promoter “Luciferase assay.”  FIG. 2  shows the results of the luciferase assay in Leiomyosarcoma tissue, primary fibroid tissue, and myometrium transfected with constructs having a Survivin promoter at 1 MOI, Survivin promoter at 5 MOI, Secretory Leukoprotease Inhibitor promoter at 1 MOI, Secretory Leukoprotease Inhibitor promoter at 5 MOI, Heparanase promoter at 1 MOI, Heparanase promoter at 5 MOI. 
     Example 3 
     In Vitro Bioluminescence Imaging 
     Materials and Methods 
     Animal Model 
     SKUT-1, 1ry F, Myo N cells (5×10 6  or 20×10 6 ) were either implanted directly or transfected with Adenovirus first then implanted into the right flank of female nude mice (Nu/Nu; Harlan Laboratory), and tumors developed over a period of 3-5 days. 
     Luciferin D In Vitro Bioluminescence Imaging 
     To confirm the superior Ad Survivin controlled reporter gene expression by IVIS live cell-imaging, the same number of cells were transfected and imaged using Xenogen IVIS 100 (Caliper Life Sciences, Hopkinton, Mass.), an optical imaging device with extremely light-tight, low background imaging chamber. The device was used for in vitro, ex vivo, and in vivo bioluminescence detection. Firefly D-Luciferin 15 mg/ml in PBS was added to cell culture media in culture dishes. The software mode was set to luminescence, photography and X-ray. Exposure was 600 with low binning (Henriques, Henriques-Pons, et al. 2014). SKUT-1 Cells are highly expressing luciferase enzyme under survivin promoter in comparison to benign tumor cells as well as healthy myometrial cells in vitro by “IVIS Live cell Bioimaging” 
     Results 
     SKUT-1 Cells highly express luciferase enzyme under survivin promoter control in comparison to benign tumor cells as well as healthy myometrial cells in vitro by “IVIS Live cell Bioimaging” ( FIG. 3 ). The order of dishes is Leiomyosarcoma, Primary Fibroid and Myometrium respectively.  FIG. 4  shows the correlation of the total photon emission/second to the MOI of the different studied cell lines. 
     Example 4 
     Ex Vivo and In Vivo Bioluminescence Imaging 
     Materials and Methods 
     Luciferin-D Ex Vivo and In Vivo Bioluminescence Imaging 
     Either ex vivo or in vivo transfection was utilized in animal studies. In the ex vivo case, transfected cells were implanted in the animals either subcutaneously or intrauterine. Untransfected cells were implanted in the nude mice and then, when the tumor developed the virus was injected intravenously. D-Luciferin 15 mg/ml in PBS was intraperitoneally injected at 260 ul per mouse followed by isofluran inhalation anesthesia 10 minutes later. The anesthetized animals were then placed in the IVIS chamber. The software mode was set to luminescence, photography and X-ray. Exposure was 600 with low binning. (Henriques, C., et al., Parasit Vectors 7: 89 (2014)). 
     Results 
     Ex vivo bioimaging of subcutaneously injected cells shows higher expression in LMS compared to benign and normal cells ( FIGS. 5A and 5B ). Ex-vivo subcutaneous model Bioluminescence imaging at 5×10 6  cells per animal thirty minutes ( FIG. 5A ) and one hour ( FIG. 5B ) post D-luciferin injection. Showing signal only in the transfected LMS lesions and is near zero in the non-transfected as well as the begin leiomyoma case. (P&lt;0.0001) 
     In vivo Bioimaging showing Leiomyosarcoma lesions emits significantly higher luminescence compared to benign and normal tissue ( FIGS. 6A-6C ). Bioluminescence imaging subcutaneously injected with 20×10 6  cells per animal transfected with Ad5-Survivin-luc. Signal shows only in the transfected LMS lesions and is near zero in the non-transfected as well as the begin leiomyoma case. (P&lt;0.0001). 
       FIGS. 7A-7B  show images of mice with 20×10 6  cells injected intracervically. The cells are t-LMS, t-1ryF, or LMS cells.  FIG. 7A  is shows images thirty minutes post injection.  FIG. 7B  shows images one hour post injection, and  FIG. 7C  shows images post cellular implantation with the same number of cells.  FIG. 7D  is line graph of total photon emission per second versus time for from top to bottom t-LMS, t-1ryF, and LMS cells. 
       FIG. 8  is an image of mice injected with 5×10 6  cells intracervically. The cells are t-LMS, t-LMS, LMS, t-1ryF, and t-1ryF cells from left to right. Signal shows only in the infected LMS lesions and is near zero in the non-transfected as well as benign leiomyoma case. 
     While in the foregoing specification this invention has been described in relation to certain embodiments thereof, and many details have been put forth for the purpose of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein can be varied considerably without departing from the basic principles of the invention. 
     All references cited herein are incorporated by reference in their entirety. The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof and, accordingly, reference should be made to the appended claims, rather than to the foregoing specification, as indicating the scope of the invention.