bluuebunny/crossref_metadata_2025 huggingface dataset:
Decreased cytotoxic effects of doxorubicin in a human ovarian cancer‐cell line expressing wild‐type p53 and WAF1/CIP1 genes
Authors: Vikhanskaya Faina , Broggini Maurizio D'Incalcimassimo
Published: February 2007

The cytotoxicity of Doxorubicin and cis‐dichloro‐diammine‐platinum (DDP) was evaluated in clones, obtained from a human ovarian cancer cell line transfected with a temperature‐sensitive p53 mutant, which express mutant p53 at 37°C and wild‐type‐like p53 at 32°C. DDP was equally active in cells not expressing p53 (SKN) or cells expressing a mutated form of p53 (SK23a kept at 37°C) or a wild‐type‐like form of p53 (SK23a cells kept at 32°C). In contrast, Doxorubicin was less cytotoxic in cells expressing wild‐type p53 than in cells expressing no p53 or mutated p53. This reduction was not due to a decreased intracellular accumulation or to a faster efflux of Doxorubicin. Topoisomerase II was found to be present in the same amount in all the systems utilized and to be functionally active, thus not accounting for the observed effect of Doxorubicin. A clear induction of WAF1 / CIP1 and GADD45 genes in cells expressing wild‐type p53 after Doxorubicin treatment was found. DDP, which was equally active in the cells utilized, caused an increase in the transcription only of GADD45 gene but not of WAF1 / CIP1 gene. Doxorubicin was also able to induce the transcription of WAF1 / CIP1 gene in SKN cells (not expressing p53) or in SK23a cells at 37°C (expressing mutated p53), indicating that the expression of this gene also, in some tumor‐cell lines, is not necessarily or uniquely induced by wild‐type p53. © 1995 Wiley‐Liss, Inc.
Escape mechanisms of human leukemic cells to long‐term immunotoxin treatment in an in vitro experimental model
Authors: Chignola Roberto , Pasti Marcella , Candiani Carola , Franceschi Antonia , Anselmi Cristina , Tridente Giuseppe , Colombatti Marco
Published: February 2007

In kinetic assays, an anti‐CD5‐ricin A chain (ST.1 ‐RTA) immunoconjugate (immunotoxins, IT) specifically inhibited up to 40% the protein synthesis of Jurkat target cells within the first 40 hr. Longer exposures of leukemia cells to ST.I‐RTA resulted in a progressively higher number of target cells escaping IT treatment and becoming resistant to further treatment with ST.I‐RTA even in the presence of the RTA‐IT enhancer monensin. Resistant Jurkat cells proliferated at the same rate as control untreated cells, and were as sensitive as control cells to a transferrin‐RTA IT, indicating that the ST.I‐RTA‐resistant tumor‐cell population did not become insensitive to the enzymatic activity of RTA. Binding studies revealed that the anti‐CD5 IT treatment induced a transient modulation of CD5 antigens but not of the functionally related CD3 antigens. The CD5 antigens were re‐expressed at the cell surface following removal of the IT molecules from the culture medium with 1.1% of the total CD5 Ag being re‐expressed per hr. When our experimental data on the kinetics of cell intoxication by the IT were corrected for the proliferative potential of the resistant and of the sensitive tumor‐cell populations, it appeared that the effect of ST.I‐RTA treatment on Jurkat cells was only to delay cell growth for a limited time period (20 hr) without reducing effectively the tumor‐cell burden. Our results may have implications for the long‐term treatment of target tumor cells with IT. © 1995 Wiley‐Liss, Inc.
Adoptive transfer of ex vivo‐activated memory T‐cell subsets with cyclophosphamide provides effective tumor‐specific chemoimmunotherapy of advanced metastatic murine melanoma and carcinoma
Authors: Gold Jay E. , Zachary David T. , Osband Michael E.
Published: February 2007

Autolymphocyte therapy (ALT) is adoptive cellular therapy of neoplastic disease using ex vivo activation of autologous (human) or syngeneic (murine) lymphocytes from tumor‐bearing hosts (TBH) by low doses of anti‐CD3 monoclonal antibody (MAb) and a mixture of previously prepared autologous cytokines (T3CS). Ex vivo activation by T3CS without tumor antigen results in expansion of CD44+ (memory) T cells. These memory T cells (ALT cells) mediate in vivo anti‐tumor specificity and with cyclophosphamide (CY) are capable of curing metastatic disease in murine TBH. To determine whether CY could enhance the effectiveness of CD4+ or CD8+ subsets of ALT cells, C57BL/6J TBH with B16 melanoma or Lewis lung (3LL) carcinoma were treated with adoptive chemoimmunotherapy (ACIT) using CD4‐depleted or CD8‐depleted ALT cells and CY. ALT cells were derived from splenocytes of B16 or 3LL‐TBH and activated ex vivo with T3CS. Depletion of CD4+ or CD8+ T cells was performed before or after activation with T3CS. B16‐TBH or 3LL‐TBH that received ACIT using CY with B16‐derived or 3LL‐derived CD8‐depleted ALT cells, respectively, demonstrated cure of metastatic disease regardless of whether CD8+ T cells were depleted before or after T3CS activation. B16 or 3LL‐TBH that received ACIT using CY with B16 or 3LL‐derived CD4‐depleted ALT cells also cured metastatic disease but only if CD4+ T cells were depleted after T3CS activation. Interleukin (IL)‐2 added to pre‐T3CS CD4‐depleted ALT cells cultured with T3CS restored anti‐tumor activity when combined with CY. TBH cured by ACIT using CY and ALT‐cell subsets derived from syngeneic TBH with the identical tumor displayed tumor‐specific immunity in rejecting a lethal challenge of identical but not reciprocal tumor. TBH given ACIT using CY and ALT‐cell subsets derived from splenocytes of syngeneic TBH with reciprocal tumors rejected lethal challenges of both tumors. Tumor specificity measured by interferon (IFN)‐γ and 51Cr‐release assays was demonstrated in pre‐ or post‐T3CS/CD8‐depleted, post‐T3CS/CD4‐depleted and pre‐T3CS + IL‐2/CD4‐depleted ALT‐cell subsets. Our data demonstrate that ACIT using CY combined with ex vivo T3CS‐activated CD44+ memory T‐cell subsets conveys long‐term tumor‐specific immunity. © 1995 Wiley‐Liss, Inc.
Quantitative variation of proto‐oncogene and cytokine gene expression in isolated breast fibroblasts
Authors: Spanakis Emmanuel , Brouty‐Boyé Danièle
Published: February 2007

Transcripts coding for transcription factors (RB, P53, FOS, MYC, MYB, ERBA, REL), growth factors (FGFI, FGF2, INT2, TGFA, TGFB, PDGF, IGF1, IGF2), interleukins, (IL1, IL2, IL3, IL4, IL6, TNF), growth‐factor receptors or cytosolic protein kinases (RAF, PIM, FES, MET, SRC, ROS, TRK, KIT, CSFR, IGFR, PDGFR, EGFR, NEU) were quantified in cultured human mammary fibroblasts from normal tissues, benign tumours, carcinomas and post‐radiation fibrosis lesions by slot‐blot auto‐radiography and image analysis. The effects of a differentiating agent (cholera toxin) and of a tumour promoter (12‐O‐tetradecanoyl‐phorbol‐13‐acetate) were also examined. The drugs modulated the levels of the anti‐oncogene transcripts (RB, P53) and of ERBA, REL, RAF, MET, ROS, TRK, CSFR, EGFR, NEU, FGF1, INT2, IGF1, IL1, IL2, IL4 and IL6. Apart from this variation, there were multiple differences in gene expression among normal and pathological cells (concerning all but P53, TGFB and interleukin transcripts) and between sub‐types defined by the presence of α‐sm‐actin (myofibroblasts) or EDB‐fibronectin (RAF, ROS, FES, KIT, IGFR, NEU, INT2, TGFB, PDGF, IGFs, ILs). It appears, therefore, that mammary stroma progress irreversibly along with the epithelium during tumoral development, and that breast cancer is not only a multi‐gene but also a multi‐tissue phenotype. © 1995 Wiley‐Liss, Inc.
High incidence of bk virus large‐T‐antigen‐coding sequences in normal human tissues and tumors of different histotypes
Authors: De Mattei Monica , Martini Fernanda , Corallini Alfredo , Gerosa Massimo , Scotlandi Katia , Carinci Paolo , Barbanti‐Brodano Giuseppe , Tognon Mauro
Published: February 2007

T‐antigen (TAg) coding sequences specific for BK virus (BKV) were detected, by PCR amplification followed by Southern‐blot hybridization, in a high percentage of human tumors and tumor‐cell lines, as well as in normal tissues, by analysis of 189 specimens. Specifically, the BKV early region was detected in 85% of brain tumors and in all normal brain tissues, in 78% of osteosarcomas, in 38% of Ewing's tumors, in 40% of normal bone specimens and in 71% of normal peripheral blood cell samples. Wilms' tumor tissues used as a control were all negative for BKV sequences. RT‐PCR analysis indicated that TAg coding sequences were expressed in specimens carrying BKV early region, ranging from 64% of the osteosarcomas to 100% of glioblastomas, Ewing's tumors, peripheral blood cells and normal bone. Moreover, DNA sequencing performed in 12 different positive samples revealed that the amplified PCR products are identical to the early‐region sequence of wild‐type BKV. The role of BKV TAg and its possible mechanism of action in human tumorigenesis are discussed. © 1995 Wiley‐Liss, Inc.
Gene transfer of human TNFα into glioblastoma cells permits modulation of mdr1 expression and potentiation of chemosensitivity
Authors: Walther Wolfgang , Stein Ulrike , Pfeil Dagmar
Published: February 2007

Despite substantial advances in the surgery, radiotherapy and chemotherapy of gliomas, the prognosis of patients with glioblastomas has still not improved. Disappointing results in chemotherapy of glioblastomas resulting from multi‐drug resistance (MDR) prompted us to investigate the influence of cytokine gene transfer in glioblastoma cells on the expression of P‐glycoprotein and on chemosensitivity of transduced cells. Several investigations have shown that malignant gliomas express P‐glycoprotein at high levels. The P‐glycoprotein is a product of the multi‐drug resistance gene (mdr 1) and functions as an energy‐dependent efflux pump which decreases drug accumulation and cytotoxicity. Since tumour necrosis factor alpha (TNFα) is a powerful anti‐cancer agent used in clinical trials and gene therapy protocols, this cytokine gene was chosen for the present investigations. Transduction of the human TNFα (hTNF) gene carrying retrovtral vector pN2tk‐hTNF into U373MG human glioblastoma cells resulted in expression and secretion of biologically active hTNF. Release of transduced hTNF reduces P‐glycoprotein expression and is associated with enhanced rhodamine‐123 uptake and potentiation of cytotoxicity of the MDR‐relevant drugs vincristine and doxorubicin. Furthermore, the transfected cell clones showed a reduced growth rate compared to the parental cells. © 1995 Wiley‐Liss, Inc.
Cellular death in neuroblastoma: in situ correlation of apoptosis and bcl‐2 expression
Authors: Hoehner Jeff C. , Hedborg Fredrik , Wiklund Helena Jernberg , Olsen Leif , Påhlman Sven
Published: February 2007

Apoptosis is the selective physiologic deletion of cells that are no longer required. Over‐expression of the bcl‐2 protooncogene extends survival of neurons otherwise destined for apoptosis. The unique capacity of neuroblastoma (NB) to undergo spontaneous regression and the prognostic dichotomy of children with this malignancy led us to evaluate bcl‐2 expression and apoptosis in NB. An in situ DNA nick‐labeling technique to detect apoptotic cells, as well as immunohistochemistry and morphology, were utilized in a selection of NB tumor specimens and in the human fetal sympathetic nervous system. bcl‐2 expression was present in all 28 NB tumors examined and in sympathetic ganglia of the human fetus. Measurement of overall bcl‐2 expression and of extent of apoptosis correlated with favorable prognosis. In low‐stage tumors, bcl‐2 expression was most intense in poorly differentiated tumor cells adjacent to fibroyascular stroma. Cells distant from the stroma exhibited increasing degrees of chromaffin differentiation, with apoptosis most evident in bcl‐2‐negative neuroblasts adjacent to well‐differentiated NB cells. The spatial distribution of bcl‐2 expression, apoptosis and chromaffin differentiation in favorable‐prognosis NB may provide insight into mechanisms of persistent tumor existence or regression. © 1995 Wiley‐Liss Inc.
Molecular events underlying schistosomiasis‐related bladder cancer
Authors: Ramchurren Nirasha , Cooper Kum , Summerhayes Ian C.
Published: February 2007

Twenty‐one invasive squamous‐cell carcinomas (SCC) of the bladder from Schistosoma‐hematobium‐infected patients were examined immunohistochemically for the expression of p53, Rb, EGFR and c‐erbB‐2 protiens; and screened by single‐strand conformation polymorphism and sequencing for mutations in the ras (H, N, K) codon hotspots (12, 13, 61) and p53 (exons 4‐9) genes. Positive staining for p53, EGFR and c‐erbB‐2 was reported in 39, 67 and 28% of tumors respectively. Only one of the tumors, the oly one that was poorly differentiated, displayed an absence of nuclear Rb staining. Ras alterations were detected in the H‐ras gene in 3 tumors, 2 of which harbored a condon‐13 (Gly → Arg) and one condon‐12 (Gly → Ser) point mutation. p53 mutations were recorded in 12 tumors (57%), 6 of which stained positively for p53. Four tumors had exon‐7 mutations (codons 235, 241 and 249; one tumor had 2 exon‐7 mutations). Eight tumors were mutated in exon 8 (codons 264, 271, 273, 285, 286, 288 and 294), 5 of which harbored miltiple mutations. One tumor had an insertion/deletion event in exon 9. The frequency of detection of over‐expression of EGFR and c‐erbB‐2 in bilharzial‐bladder lesions is comparable to that reported in TCC, contrasting with the infrequent loss of Rb expression found in invasive lesions associated with schistosomiasis infection. However, the detection of multiple p53 mutations in these lesions is suggestive of the involvement of a carcinogenic agent with maintenance of preferential activation of the H‐ras gene. © 1995 Wiley‐Liss Inc.
In situ expression of B7 and CD28 receptor families in human malignant melanoma: Relevance for T‐cell‐mediated anti‐tumor immunity
Authors: Denfeld Ralf W. , Dietrich Andrea , Wuttig Christiane , Tanczos Eszter , Weiss Johannes M. , Vanscheidt Wolfgang , Schöpf Erwin , Simon Jan C.
Published: February 2007

Work in animal models has suggested that interactions of members of the B7 receptor family (e.g., B7–1, B7–2) on tumor cells with their ligands CD28 and CTLA‐4 on cytotoxic T cells (CTL) are important for the induction of anti‐tumor immunity against malignant melanoma (MM). To determine whether these molecules are of relevance for CTL responses against human MM, we studied the expression of B7–1, B7–2, CD28 and CTLA‐4 in primary tumors of MM (PMM), MM metastases (MMM) and benign melanocytic nevi (BMN) by immunohisto‐chemistry (IH) and by reverse transcription polymerase chain reaction (RT‐PCR). By RT‐PCR, B7–1 and B7–2‐specific mRNAs were detected in most PMM, MMM and BMN. These PCR‐signals were derived from CD45+‐infiltrating leukocytes and not from tumor cells since (1) MMM depleted of CD45+ cells contained no B7–1 or B7–2 mRNA; and (2) by IH, B7–1 and B7–2 were found on infiltrating dendritic cells, macrophages and a variable proportion of tumor‐infiltrating lymphocytes (TIL) but not on melanoma cells or nevus cells. The important exceptions were 5/5 spontaneously regressing PMM, in which B7–1 and B7–2 were expressed by melanoma cells, that were surrounded by TIL expressing CD28 but not CTLA‐4. We conclude that, in PMM, MMM and BMN, the majority of TIL are CD28+ and that B7–1 and B7–2 are expressed by CD45+‐infiltrating antigen‐presenting cells (APC) and TIL, but not by the tumor cells. However, in spontaneously regressing PMM, melanoma cells express B7–1, B7–2 and MHC class‐I and‐II antigens, particularly in areas with clinical and histological signs of an ongoing anti‐tumor response. These data suggest that the absence of B7–1 and B7–2 favors the escape of MM from immunosurveillance, while B7–1, B7–2 expression enhances T‐cell‐mediated anti‐tumor immunity. © 1995 Wiley‐Liss Inc.
Risk of cutaneous melanoma associated with pigmentation characteristics and freckling: Systematic overview of 10 case‐control studies
Authors: Bliss Judith M. , Ford Deborah , Swerdlow Anthony J. , Armstrong Bruce K. , Cristofolini Mario , Elwood J. Mark , Green Adèle , Holly Elizabeth A. , Mack Thomas , Mackie Rona M. , ØSterlind Anne , Walter Stephen D. , Peto Julian , Easton Douglas F.
Published: February 2007

Using individual subject data from 10 case‐control studies, comprising over 3000 cases and almost 4000 controls, we have estimated the relative risk of melanoma associated with aspects of complexion, namely, hair, eye and skin colour and freckling in adulthood, and have examined the relationships between these factors and naevus count in terms of melanoma risk. Compared with individuals with black or dark brown hair, the relative risks for developing melanoma in those with light brown, blonde and red hair were 1.49 (95% Cl 1.31, 1.70), 1.84 (95% Cl 1.54, 2.21) and 2.38 (95% Cl 1.90, 2.97), respectively. Individuals with blue eyes had a risk 1.55 (95% Cl 1.35, 1.78) times that for those with brown eyes, or 1. 15 (95% Cl 0.94, 1.40) after adjusting for hair colour and freckling in adulthood. The relative risks associated with hair and eye colour were independent of those for naevus count and skin colour. Light skin colour and high freckle density were also highly significant risk factors, independent of each other and of naevus count and hair and eye colour. The risks associated with these factors, while individually modest, are largely independent, and thus pigmentation characteristics and freckling tendency should be useful in identifying high risk groups to be targeted for prevention. © 1995 Wiley‐Liss, Inc.
Detection and characterization of the prostate‐specific membrane antigen (PSMA) in tissue extracts and body fluids
Authors: Trover John K. , Beckett Mary Lou , Wright George L.
Published: February 2007

The prostate‐specific membrane antigen (PSMA) glycoprotein is recognized by the monoclonal antibody (MAb) 7EII ‐C5.3 as a predominant 100 kDa and minor 180 kDa component in LNCaP cell line extracts and its expression has been shown by immunohistochemistry to be highly restricted to prostate epithelium. The aim of the present study was to utilize Western blot analysis to determine if PSMA could be detected in human tissue extracts and body fluids and if so, which molecular forms were present. PSMA was detected as 120 and 200 kDa bands in normal, benign and malignant prostate tissues and seminal plasma. Further analysis demonstrated that the larger molecular form of PSMA may be a dimer of the lower m.w. species. The PSMA glycoprotein was not detected in the majority of non‐ prostate tissue extracts examined except for a low yet significant amount in normal salivary gland, brain and small intestine, suggesting that PSMA may not be as prostate‐specific as originally thought. Since the prostate‐specific antigen (PSA) has been shown to be maximally shed into the serum in high‐grade and metastatic prostate carcinomas, it was surprising that PSMA could not be detected in serum by Western blot analysis even in patients with actively progressive metastatic disease. Second generation antibodies generated against different epitopes may be required to determine if PSMA is shed into serum. Our results support the hypothesis that PSMA is a novel prostate biomarker. © 1995 Wiley‐Liss, Inc.
Reproductive and other factors and risk of epithelial ovarian cancer: An australian case‐control study
Authors: Purdie David , Green Adèle , Bain Christopher , Siskind Victor , Ward Bruce , Hacker Neville , Quinn Michael , Wright Gordon , Russell Peter , Susil Beatrice
Published: February 2007

Of the few factors known to be associated with epithelial ovarian cancer, the most consistently observed relate to women's reproductive function, although even here uncertainties remain. We have undertaken a case‐control study involving personal interviews with over 1,600 women, the largest of its kind to date, to investigate further the associations between women's reproductive histories and other factors and the development of ovarian cancer. Cases were drawn from women diagnosed with epithelial ovarian cancer in 3 Australian states, Queensland, New South Wales and Victoria, between August 1990 and December 1993, and controls were drawn at random from the electoral roll, stratified by age and geographic region. Trained interviewers administered standard questionnaires to obtain detailed information about women's reproductive and contraceptive histories and other factors of interest, such as smoking and family history of ovarian or other cancer. Findings were based on data from 824 cases and 860 controls and confirmed the reduced risk of ovarian cancer associated with increasing parity and duration of use of the oral contraceptive pill (OCP), hysterectomy and tubal ligation. The strongest association of all was seen with use of the OCP for 10 years or more. An inverse association between ovarian cancer and age at first birth was observed, but this was not statistically significant. There were no associations between development of ovarian cancer and number of incomplete pregnancies, use of hormone replacement therapy or menstrual history. Among other factors considered, education after leaving school was negatively associated and high body mass index, family history of ovarian cancer, use of talc in the abdominal or perineal region and smoking were positively associated with occurrence of ovarian cancer. © 1995 Wiley‐Liss, Inc.
Expression of urokinase receptor in various stromal‐cell populations in human colon cancer: Immunoelectron microscopical analysis
Authors: Ohtani Haruo , Pyke Charles , Dan Ø Keld , Nagura Hiroshi
Published: February 2007

The host reaction is an important factor in the biological behavior of cancers. In human colon adenocarcinoma, stromal cells and some cancer cells express the urokinase receptor (uPAR), a molecule involved in the regulation of extracellular proteolysis. The present study reveals the identity of uPAR‐ expressing cell types and the subcellular localization of this molecule by immunoelectron microscopy in colon cancer. uPAR‐positive cells were most abundant at the invasive margin of colon cancer and were identified as macrophages, fibroblasts, neutrophilic and eosinophilic granulocytes, endothelial cells and cancer cells. Of these, the most numerous were macrophages with uPAR detected along the plasma membrane, in accordance with its function in plasminogen activation on the cell surface. Fibroblasts were labeled in the lumen of rough endoplas‐ mic reticulum, indicating its intracellular synthesis. Some granulocytes and endothelial cells expressed immunoreactivity along the plasma membranes. uPAR‐positive cancer cells were stained along the plasma membrane and in rough endoplasmic reticulum. These findings suggested that a variety of non‐malignant host cells play an important role in the plasmin‐mediated breakdown of the extracellular matrix at the invasive margin. © 1995 Wiley‐Liss, Inc.
in vivo effects of 4‐amidinoindan‐l‐one 2′‐amidinohydrazone (CGP 48664A) and α‐difluoromethylornithine (DFMO) on L1210 growth, cell‐cycle phase distribution and polyamine contents
Authors: Dorhout Bernard , Te Velde Roelf Jakob , Ferwerda Harri , Kingma Anneke W. , de Hoog Elly , Muskiet Frits A. J.
Published: February 2007

We studied the in vivo effects of 4‐amidinoindan‐l‐one 2′‐amidinohydrazone (CGP 48664A), α‐difluoromethylornithine (DFMO) and a combination of CGP 48664A‐DFMO on tumor growth, cell‐cycle phase distribution and polyamine contents. DBA‐2 mice were inoculated i.p. with 105 L1210 cells on day 0, treated i.p. on days 1–4 and killed on day 5. As compared to controls, CGP 48664A, DFMO and the CGP 48664A‐DFMO combination reduced L1210 cell numbers by 33, 43 and 85%, respectively. CGP 48664A did not affect cell‐cycle phase distribution. DFMO and the CGP 48664A‐DFMO combination caused a moderate and a heavy accumulation in G0/G1‐ and G2/M‐phases, respectively. Compared with controls, the CGP 48664A‐DFMO combination reduced putrescine, spermidine and total polyamines, but did not affect spermine. Compared with CGP 48664A, the CGP 48664A‐DFMO combination caused lower putrescine and total polyamines, higher spermine, but no change in spermidine. Compared with DFMO, the CGP 48664A‐DFMO combination caused higher putrescine and spermidine, lower spermine, but no change in total polyamine levels. We conclude that CGP 48664A potentiates the cystostatic effect of DFMO in vivo. The resulting growth inhibition is accompanied by an accumulation in G0/G1‐ and G2/M‐phases and a reduction of putrescine and spermidine. The data suggest that perturbed polyamine composition rather than reduced spermidine or total polyamine pool size causes a profound growth inhibition. © 1995 Wiley‐Liss, Inc.
Cyclophosphamide plus tumor necrosis factor‐α chemoimmunotherapy cured mice: Life‐long immunity and rejection of re‐implanted primary lymphoma
Authors: Ehrke M. Jane , Verstovšek Srdan , Krawczyk Christian M. , Ujházy Peter , Zaleskis Gintaras , Maccubbin Darbie L. , Mihich Enrico
Published: February 2007

Changes in functionally and phenotypically definable spleno‐cyte subsets in aging mice which had been rendered tumor‐free in early life by immunochemotherapy (cyclophosphamide plus tumor necrosis factor‐α) were studied in the syngeneic EL4 lymphoma‐C57BL/6 murine model. Treatment‐induced long‐term survivors (LTS) surviving rechallenge are termed “immune‐LTS”. On day 120 (day 0, initial tumor inoculation), splenocytes from day 60 rechallenged immune‐LTS developed significantly greater specific anti‐EL4 cytolytic activity in an ex vitro assay than those from non‐rechallenged LTS. Splenocytes from combination‐treated groups developed significantly higher activity than those from cyclophosphamide‐induced immune‐LTS. The splenic effector precursor was a CD8+ T cell. The specific anti‐EL4 effector cell from the cyclophosphamide‐induced immune‐LTS was CD4− CD8+; however, approximately 50% of those from combination‐treated immune‐LTS appeared to be CD4+CD8+. On day 520 immune‐LTS were randomized into 2 groups. One group was re‐implanted with EL4 tumor; all mice survived. The other group was killed and, even though their splenocytes developed considerable anti‐EL4 activity, their allogeneic responsiveness was as reduced as that of age* matched controls. Phenotypic analysis, compared with splenocytes from young and age‐matched controls, revealed changes in the makeup of each T‐cell subset, except the CD4+CD8+, and all subsets, except the CD4− CD8−, had increases in CD44 positivity. On day 625, the age of these mice was equivalent to the median life‐span of C57BL/6 mice; nevertheless, their splenocytes developed high anti‐EL4 activity. Phenotypic analysis indicated that, compared to day 520, there was a major decrease in CD4− CD8+ splenocytes; we suggest that these cells had migrated to the site of tumor eradication.
EWS‐FLI‐1 and EWS‐ERG chimeric mRNAs in Ewing's sarcoma and primitive neuroectodermal tumor
Authors: Ida Kohmei , Kobayashi Shigetoshi , Taki Tomohiko , Hanada Ryoji , Bessho Fumio , Yamamori Shunji , Sugimoto Tohru , Ohki Misao , Hayashi Yasuhide
Published: February 2007

Thde t( 11;22)(q24;q 12) and t(21;22)(q22;q 12) are specific chromosomal translocations found in the Ewinhd family of tumors including ES, PNET and Askin tumors. In these translocations, the amino‐terminal portion of the EWS gene located in 22q 12 fuses to the carboxyl‐terminal portion of the FLI‐I gene located in 11 q24 or the ERG gene located in 21 q22, which belong to the ets oncogene super‐family of transcription activators. We investigated the chimeric mRNAs of 15 ESs (7 cell lines and 8 tumor samples) and 7 PNETs (3 cell lines and 4 tumor samples) using the RT‐PCR method and sequencing. We detected 2 types of EWS‐ERG chimeric mRNA in 2 ES cell lines and I PNET tumor sample in addition to 4 types of EWS‐FU‐1 chimeric mRNA in 11 ESs (4 cell lines and 7 tumor samples) and 4 PNETs (2 cell lines and 2 tumor samples). There seemed to be no association between the type of chimeric mRNA and clinical features such as sex, age, primary site and histopathology of the patients. All of the chimeric mRNAs are generated from in‐frame junctions and are thought to encode fusion proteins that may be the molecular mechanism involved in the Ewing family of tumors.
Specific targeting of a mutant, activated egf receptor found in glioblastoma using a monoclonal antibody
Authors: Hills David , Rowlinson‐Busza Gail , Gullick William J.
Published: February 2007

A truncated epidermal growth factor receptor (EGFR) expressed from a rearranged and amplified EGFR gene is present at high frequency in gliomas. In this work we show that when this receptor is expressed in NIH3T3 fibroblasts it is partially activated and confers tumorigenicity to this cell line in vivo but no growth advantage in vitro anchorage‐independent growth assays. Because the mutation occurs in the extracellular domain of the receptor, it can be considered to represent a glioma‐specific tumour marker. Here we demonstrate that 2 monoclonal antibodies, DH 1.1 and DH8.3, raised to a synthetic peptide spanning the unique junctional sequence, can recognise the mutant receptor but not the normal receptor in both denatured and native states. Furthermore, radiolabelled antibody DH8.3 successfully targets tumours expressing this antigen in nude mice.
Prevention of prostate‐cancer metastasis in vivo by a novel synthetic inhibitor of urokinase‐type plasminogen activator (uPA)
Authors: Rabbani Shafaat , Harakidas Penelope , Davidson Donald J. , Henkin Jack , Mazar Andrew P.
Published: February 2007

Urokinase‐type plasminogen activator (uPA) is a serine protease associated with tissue remodeling, cellular invasiveness, matrix degradation and tumor growth. Over‐expression of uPA by the rat prostate‐cancer cell line Dunning R3227, Mat LyLu, results in increased tumor metastasis to several non‐skeletal and skeletal sites. Histological examination of these skeletal lesions has shown them to be primarily osteoblastic. In the present study we examined the capacity of a selective inhibitor of uPA enzymatic activity, 4–iodo benzo[b]thiophene‐2–carbox‐amidine (B‐428), to prevent the development of tumor growth and invasiveness in a syngeneic model of rat prostate cancer using a Dunning R3227 cell line over‐expressing rat uPA. Male Copenhagen rats were inoculated s.c. with experimental cells into the right flank and continuously infused i.p. with either vehicle alone or uPA inhibitor for 2 to 3 weeks. Animals were killed at timed intervals and evaluated for the development of tumor growth and metastasis. Serum from these animals was collected to examine any signs of nephrotoxicity. Control animals receiving vehicle alone developed large tumors at the site of inoculation as well as macroscopic metastases in the lungs, kidney, spleen and lymph nodes. In contrast, experimental animals receiving uPA inhibitor showed a marked decrease in primary tumor volume and weight as well as in the development of tumor metastases. The occasional tumor metastases observed after infusion of B‐428 were significantly smaller than those observed in vehicle controls. These effects of B‐428 were found to be dose‐dependent without any adverse effects on the renal function of experimental animals. These studies demonstrate that uPA‐specific inhibitors can decrease primary tumor volume and invasiveness as well as metastasis in a model of prostate cancer where uPA has been implicated as a major pathogenetic factor. © 1995 Wiley‐Liss, Inc.
Allelic loss at 1p and 19q frequently occurs in association and may represent early oncogenic events in oligodendroglial tumors
Authors: Josefa Bello M. , Leone Paola E. , Vaquero Jesús , De Campos José M. , Elena Kusak M. , Sarasa José L. , Pestaña Angel , Rey Juan A.
Published: February 2007

The molecular mechanisms underlying the genesis and progression of oligodendroglial tumors are poorly understood, since only restricted information on loss of heterozygosity from isolated cases is available. The commonest alterations appear to involve deletion of 1p and 19q, while loss of heterozygosity for 9p, chromosome 10 or epidermal growth factor receptor gene amplification have been described in single tumors. We have applied restriction fragment length polymorphism analysis to 14 loci covering chromosome 1 and 7 loci on chromosome 19 in a series of 25 tumors with an oligodendroglial component to determine precisely the participation of these suppressor genes in the genesis of tumors. Twenty‐two and 19 of the 25 samples displayed LOH at 1p and 19q, respectively, and both anomalies were detected in association in 17 samples, including low‐ and high‐grade oligodendrogliomas as well as mixed oligo‐astrocytomas. Our findings suggest that inactivation of tumor suppressor genes located on 1p and 19q represent cooperative alterations occurring at early stages of oncogenic transformation of oligodendroglial neoplasms. © 1995 Wiley‐Liss, Inc.
E‐cadherin expression in bladder cancer using formalin‐fixed, paraffin‐embedded tissues: Correlation with histopathological grade, tumour stage and survival
Authors: Syrigos Konstantinos N. , Krausz Thomas , Waxman Jonathan , Pandha Hardev , Rowlinson‐Busza Gail , Verne Julia , Epenetos Agamemnon A. , Pignatelli Massimo
Published: February 2007

To determine the potential prognostic value of epithelial cadherin (E‐cadherin), a Ca2+‐dependent cell‐cell adhesion molecule, we have analysed its immunoreactivity and cellular localisation in 67 transitional cell carcinomas (TCC) using an avidin‐biotin immunoperoxidase technique on formalin‐fixed, paraffin‐embedded tissues. These results were correlated with histopathological grade, tumour stage, presence of metastases and survival. In addition, 10 cystitis and 11 normal bladder biopsies were evaluated as controls. E‐cadherin was expressed in a normal membranous pattern in all normal and 7 of 10 cystitis biopsies. Loss of normal surface E‐cadherin was expression was found in 3 of 15 superficial tumours and in 48 of 52 invasive cancers. Abnormal immunoreactivity was strictly related to tumour differentiation and stage. Fifteen of 20 well‐differentiated (grade 1) tumours showed preserved membranous E‐cadherin immunoreactivity, while 46 of 47 moderate and poorly differentiated tumours (grades II and III) demonstrated abnormal staining patterns. Loss of membranous E‐cadherin immuno‐reactivity was also associated with advanced tumour stage. There was a significantly higher 5‐year survival rate for patients with preserved membranous staining compared with patients with abnormal staining. © 1995 Wiley‐Liss, Inc.
In vitro differentiation of HT‐29 M6 mucus‐secreting colon cancer cells involves a trychostatin A and p27KIP1‐inducible transcriptional program of gene expression
Authors: Mayo Clara , Lloreta Josep , Real Francisco X. , Mayol Xavier
Published: February 2007

Tumor cell dedifferentiation—such as the loss of cell‐to‐cell adhesion in epithelial tumors—is associated with tumor progression. To better understand the mechanisms that maintain carcinoma cells in a differentiated state, we have dissected in vitro differentiation pathways in the mucus‐secretor HT‐29 M6 colon cancer cell line, which spontaneously differentiates in postconfluent cultures. By lowering the extracellular calcium concentration to levels that prevent intercellular adhesion and epithelial polarization, our results reveal that differentiation is calcium‐dependent and involves: (i) a process of cell cycle exit to G0 and (ii) the induction of a transcriptional program of differentiation gene expression (i.e., mucins MUC1 and MUC5AC, and the apical membrane peptidase DPPIV). In calcium‐deprived, non‐differentiated postconfluent cultures, differentiation gene promoters are repressed by a trichostatin A (TSA)‐sensitive mechanism, indicating that loss of gene expression by dedifferentiation is driven by histone deacetylases (HDAC). Since TSA treatment or extracellular calcium restoration allow gene promoter activation to similar levels, we suggest that induction of differentiation is one mechanism of HDAC inhibitor antitumor action. Moreover, transcriptional de‐repression can also be induced in non‐differentiating culture conditions by overexpressing the cyclin‐dependent kinase inhibitor p27KIP1, which is normally induced during spontaneous differentiation. Since p27KIP1 downregulation in colon cancer is associated with poor prognosis independently of tumor cell division rates, we propose that p27 KIP1 may prevent tumor progression by, at least in part, enhancing the expression of some differentiation genes. Therefore, the HT‐29 M6 model allows the identification of some basic mechanisms of cancer cell differentiation control, so far revealing HDAC and p27KIP1 as key regulatory factors of differentiation gene expression. J. Cell. Physiol. 212: 42–50, 2007. © 2007 Wiley‐Liss, Inc.
Transgenic mice expressing LHX3 transcription factor isoforms in the pituitary: Effects on the gonadotrope axis and sex‐specific reproductive disease
Authors: Savage Jesse J. , Mullen Rachel D. , Sloop Kyle W. , Colvin Stephanie C. , Camper Sally A. , Franklin Craig L. , Rhodes Simon J.
Published: February 2007

The LHX3 transcription factor plays critical roles in pituitary and nervous system development. Mutations in the human LHX3 gene cause severe hormone deficiency diseases. The gene produces two mRNAs which can be translated to three protein isoforms. The LHX3a protein contains a central region with LIM domains and a homeodomain, and a carboxyl terminus with the major transactivation domain. LHX3b is identical to LHX3a except that it has a different amino terminus. M2‐LHX3 lacks the amino terminus and LIM domains of LHX3a/b. In vitro experiments have demonstrated these three proteins have different biochemical and gene regulatory properties. Here, to investigate the effects of overexpression of LHX3 in vivo, the alpha glycoprotein subunit (αGSU) promoter was used to produce LHX3a, LHX3b, and M2‐LHX3 in the pituitary glands of transgenic mice. Alpha GSU‐beta galactosidase animals were generated as controls. Male αGSU‐LHX3a and αGSU‐LHX3b mice are infertile and die at a young age as a result of complications associated with obstructive uropathy including uremia. These animals have a reduced number of pituitary gonadotrope cells, low circulating gonadotropins, and possible sex hormone imbalance. Female αGSU‐LHX3a and αGSU‐LHX3b transgenic mice are viable but have reduced fertility. By contrast, αGSU‐M2‐LHX3 mice and control mice expressing beta galactosidase are reproductively unaffected. These overexpression studies provide insights into the properties of LHX3 during pituitary development and highlight the importance of this factor in reproductive physiology. J. Cell. Physiol. 212: 105–117, 2007. © 2007 Wiley‐Liss, Inc.
Renal cell carcinoma—a review of 256 cases
Authors: Wagle Datta G. , Scal David R.
Published: February 2007

Renal cell carcinoma represents nearly 90% of all malignant tumors of the kidney. The varied aspects of this tumor's pathophysiology have been a continuing challenge to early diagnosis and successful therapy. For inapparent reasons it has been noted in this and other studies that renal cell carcinoma is twice as common in males as in females. As we have shown, renal cell carcinoma occurs in all age groups but is most common in the sixth and seventh decades. In spite of an occasional case, heredity probably does not apparently play a role in the pathogenesis of this disease. This study has shown that because of the multiple pathways of dissemination, renal cell carcinoma can be unpredictable in the sites of metastasis. The lung, however, is consistently the most common site of metastatic growth. Many experts have attempted to introduce a system of classification related to prognosis. The present experience using the classification of Flocks and Kadesky showed there was a notable drop in survival in stages III and IV as compared to I and II. Also, there was little difference in survival between patients having stage III and stage IV disease. In general, the survival statistics agree with those of others. The classic triad of renal cell carcinoma was associated with 5‐year survival in only four cases, indicating it to be a sign of advanced disease. Radical nephrectomy continues to be the initial treatment in most centers. The most promising of the newer methods is hormonal therapy, which is based on the theory that hormones underlie the mechanism of host resistance to renal cell carcinoma. There have been four satisfactory responses in 26 treated cases of renal tumor with distant metastases. However, it is still too early to make valid statements about this modality.
The anemia of chronic disorders: Experimental abscess, spleen transplantation, and red cell iron reutilization
Authors: Zacharia L. A. , Rigby P. G.
Published: February 2007

Studies of Fe59 incorporation into blood, liver, and spleens utilizing ‘cooked’ red blood cells labeled with 59‐ferrous citrate were undertaken to evaluate iron reutilization. Intraperitoneal transplantation of intact spleens was performed from mice both with and without turpentine abscess to similarly prepared mice. Presence or absence of abscess in the recipient mouse determined ‘compartmentalization’ of reutilizable iron between the recipient reticuloendothelial system (spleen and liver) and blood. Reduced iron reutilization induced by chronic inflammation seems to be predominantly due to alteration of reticuloendothelial system function as measured by Fe59 incorporation into blood from labeled ‘cooked’ red blood cells. The model studied shows that chronic inflammation is sufficient systemic cause for reduced iron reutilization and thereby a major contributor to the anemia of chronic diseases.
The effect of 5‐fluorouracil on small bowel mucosa
Authors: White Richard S. , Fuqua William B. , Barrett O'Neill
Published: February 2007

Pre‐ and post‐therapy small bowel biopsies were obtained in 19 of 20 patients who received 5‐fluorouracil for treatment of their malignant disease. Three parameters were evaluated: clinical response to therapy, which occurred in five cases (26%); development of gastrointestinal toxicity, which occurred in 13 (68%); and histopathologic change of small bowel mucosa, which was noted in 13 (68%). There was no correlation between clinical response to therapy and the development of toxicity, nor was there any between toxicity and histopathologic change in small bowel mucosa. Definite histopathologic change in each of the patients who responded to therapy was the only consistent relationship noted, although the appearance of such changes did not necessarily indicate that tumor response could be expected.
Inhibition of lymphocyte blastogenesis by platinum compounds
Authors: Khan Amanullah , Hill Joseph M.
Published: February 2007

The effect of platinum compounds cis‐platinum (II) diamminodichloride and cis‐platinum (IV) diamminotetrachloride on the phytohemagglutinin‐induced blastogenesis in lymphocytes was studied. Both compounds inhibited blastogenesis, suggesting that they may be immunosuppressive.
The value of chemotherapy in the treatment of testicular dysembryoplastic tumors
Authors: Pommatau E. , Brunat M. , Chassard J. L. , Mayer M.
Published: February 2007

Chemotherapy appears to be of help in treating dysembryoplastic testicular tumors. It seems better to treat first by surgery (lymphadenectomy) if possible, then (a) if no nodes are involved or only one has been discovered by microscope examination, with chemotherapy for one year; or (b) if two or more nodes are involved, with irradiation of the nodes and chemotherapy.

Even if lymphadenectomy is not possible, one course of chemotherapy followed by cobalt and more chemotherapy can bring results in some cases.
The regional lymph node question. To treat or not to treat: The pros and cons
Authors: Futrell J. William , Hoopes John E.
Published: February 2007

The question of whether or not to extend treatment to the regional lymph nodes when treating malignant disease remains unanswered. The clinician must individually approaise the pros and cons and derive a satisfactory treatment plan for each patient. Six reasons which are generally stated as indications for regional lymphadenectomy and seven potential arguments against lymphadenectomy are presented.
A new classification of generalized carcinoma of the breast based on response to therapy
Authors: Razis Dennis V.
Published: February 2007

Classifications of generalized carcinoma of the breast were based up to now on factors of definitely proven prognostic significance. These classifications are very useful in defining prognosis, but they help only to a limited extent in selecting treatment. We are proposing a new classification of three groups for generalized carcinoma of the breast according to objective response to therapy: to (a) responders, (b) nonresponders, and (c) patients with static disease. It is pointed out that in some cases there is discrepancy between prognosis and response to therapy, as some patients with “poor prognosis” respond dramatically to treatment while others with “good prognosis” do not respond to any type of treatment and treatment is actually harmful. It is also pointed out that our classificaiton has a dynamic meaning, as a patient can switch from one group to another. The proposed classification might be of more help in deciding the therapeutic management.
The influence of facial‐nerve sacrifice in surgery of malignant parotid tumors
Authors: Hodgkinson Darryl J. , Woods John E.
Published: February 2007

Malignant parotid tumors in 224 patients were dealt with surgically at the Mayo Clinic between 1940 and 1969. The clinical histories, including follow‐up of 5 years or more in every case, were reviewed with particular attention to sacrifice of the facial nerve in conjunction with the excision of tumor. This was done in 65 cases. High‐grade malignancy necessitated nerve sacrifice; other frequent indications were preoperative facial‐nerve paralysis, pain, and rapid growth of the tumor. Over the 30‐year review period, the trend was toward more radical surgery, often including nerve sacrifice, and away from local excision and radiotherapy. Nerve grafting or anastomosis of a sacrificed nerve was seldom performed in an attempt to lessen the cosmetic defect of facial paralysis. Later reconstructive procedures are favored.
Enhancement of hepatic and renal tumorigenesis in thyroidectomized NZR/Gd rats treated with dimethylnitrosamine
Authors: Noronha R. F. X. , Goodall C. M.
Published: February 2007

Endocrine modulation of DMN carcinogenesis was studied in NZR/Gd rats preconditioned by starving 48 hr and then injected i.p. once with 20 mg DMN/kg. Intact rats developed kidney tumors (44% TBA), most of tubular epithelial type resembling human tumors rather than mesenchymal. Thyroidectomy (Tx) 45 days before DMN significantly enhanced DMN carcinogenesis, renal carcinoma incidence increasing to 69%. Renal carcinomas showed more signs of malignancy in Tx rats. Other neoplastic responses useful for further studies included tumors in nasal epithelium (13%), liver (18%, increased to 59% in Tx rats), and lung (40%): these tumors were rare or not previously reported in single‐dose experiments in other rat strains. A sex difference in lung tumor incidence (male 70%, female 16%) was statistically significant and thyroidectomy reduced the sex‐differential (to 54% and 39% respectively). The increased incidence of kidney and liver tumors could be due to altered metabolism of DMN in tissues of Tx rats.
The establishment of a cancer center
Authors: Hutchinson William B.
Published: February 2007

This article describes the designated comprehensive cancer center in the Pacific Northwest section of the United States. The local financial, political, and medical problems associated with the initial establishment of this center are discussed. The general philosophy of the approach to cancer, cancer research, and care are included. The programs described differ very little from other similar centers with exceptions that are noted. These are the establishment of the major clinical facilities or units outside of the center proper. The utilization, not duplication, of already existing surgical, irradiation, and hospital facilities representing a departure from the usual custom is discussed. Active participation with real input by the physicians of our region through the extramural council is described.