bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2340: Ribonucleotide reductase subunit M2 promotes cell invasiveness and motility in lung and head and neck cancer cells
Authors: Rahman Mohammad Aminur , Amin A.R.M. Ruhul , Chen Zhuo Georgia , Davis Mark E. , Shin Dong Moon
Published: June 2012

Ribonucleotide reductase subunit M2 (RRM2) has been frequently observed to be aberrantly overexpressed in various tumors. RRM2 is a key enzyme involved in the production of 2′-deoxyribonucleoside 5′-triphosphates required for DNA synthesis. In addition to DNA synthesis, it has been reported that RRM2 modulates cellular invasiveness. However, the mechanisms through which RRM2 affects the invasive phenotype have not been elucidated. In our study, we examined the role of RRM2 in cell invasion and migration of different lung (A549, H460 and H1299) and head and neck (Tu686) cancer cell lines. To explore the mechanism underlying RRM2's activity, we applied small interference RNA (siRNA) to knockdown the RRM2 protein while pcDNA3/RRM2 plasmid was used to establish RRM2 overexpressing stable cell lines. Our results revealed varying degrees of RRM2 expression levels in lung and head and neck cancer cell lines, which were proportionally correlated with the invasive capability of cells. H1299, a cell line that features epithelial-mesenchymal transition (EMT) expressed the highest level of RRM2 protein among the cell lines studied, and exhibited the greatest invasive and migratory abilities as demonstrated by Boyden chamber invasion and wound healing assays. H1299 was 6-fold more invasive than A549, which has lower RRM2 expression. Moreover, gain of function analysis demonstrated that overexpression of RRM2 enhanced cell invasiveness and motility in low RRM2-expressing cell lines A549 and Tu686. Western blots revealed that markers associated with tumor cell invasion and migration such as phospho-Src, phospho-FAK, phospho-calveolin-1, phospho-Akt, phospho-Erk, and MMP-9 secretion were induced whereas E-cadherin protein expression was significantly inhibited in RRM2 overexpressing cell lines. In contrast, loss of function analysis showed that siRNA targeting RRM2 decreased RRM2 expression and attenuated cellular invasiveness and motility in both lung and head and cancer cell lines. Our study demonstrated that RRM2 substantially promoted cell invasion and migration abilities. An elevated RRM2 may serve as a potential biomarker for metastasis of head and neck and lung cancer. Thus, targeting RRM2 and its downstream signaling intermediaries represents a rational approach for developing novel anticancer therapeutics. (This work is supported by grants from NIH U54CA119338-04, P50CA128613 and DOD W81XWH-07-1-0306).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2340. doi:10.1158/1538-7445.AM2011-2340
Abstract 2354: Characterization of three novel splice variants of the RECK tumor and metastasis suppressor gene
Authors: Trombetta-Lima Marina , Winnischofer Sheila M.B. , Carreira Ana C.O. , Demasi Marcos A.A. , Colin Christian , Oba-Shinjo Sueli M. , Marie Suely K.N. , Sogayar Mari C.
Published: June 2012

REversion-inducing Cysteine-rich protein with Kazal motifs (RECK) encodes a membrane-anchored protein which suppresses both invasion and metastasis by negatively regulating at least three MMP′s, namely: MMP-9, MMP-2 and MT1-MMP. Matrix metalloproteinase (MMP) family members belong to the group of Zn2+ dependent proteolytic enzymes, which are key elements responsible for extracellular matrix (MEC) collapse, being, therefore, directly involved in tumor invasiveness and metastasis. MEC degradation by MMPs not only facilitates tumor invasion, but, also, affects the behavior of tumor and adjacent cells, leading to cancer progression. In general, the relative levels of MMPs increase with tumor progression. Other RECK targets have been identified, such as the extracellular metalloproteinases ADAM10 and CD13/ aminopeptidase N. A positive correlation has been observed between the abundance of RECK expression in tumor samples and better prognosis for patients with gastric, lung, pancreatic and colorectal cancers. Recent studies showed mutations in splicing cis regulatory elements and alterations in the cellular splicing regulatory machinery leading to changes in the splicing pattern of several cancer-related genes, such as CD44, BRCA1, APC, p53 and FHT. In the present study, three novel alternative isoforms of the RECK tumor suppressor gene, namely RECK B (1,548b), RECK D (1,737b) and RECK I (1,101b) were isolated by RT-PCR and their expression profiles were investigated using quantitative real time RT-PCR assays in a normal tissue RNA panel and during human glioma progression. Our results show that RECK isoforms display independent expression patterns when compared to the canonical form, indicating a more complex role for both the canonical and the alternative RECK isoforms in glioma progression. Support: FAPESP, CNPq, FINEP, BNDES, MCT, MS

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2354. doi:10.1158/1538-7445.AM2011-2354
Abstract 2356: Investigating the role of mammalian RNA binding protein Musashi-2 (MSI2) in breast cancer invasion and nodal metastasis
Authors: Holzapfel Nick , Nair Ranju , Done Susan J.
Published: June 2012

Previous array comparative genomic hybridization (aCGH) studies in our lab have shown that chromosomal gains on 17q22-24.2 are associated with invasion when detected in duct carcinoma in situ (DCIS) and nodal metastasis when detected in invasive duct carcinoma (IDC). Of nine candidate genes identified within this region, MSI2 was selected for further study. MSI2 is a Musashi family RNA binding protein that translationally regulates target mRNAs. RNAi screens have indicated that MSI2 down regulation impairs hematopoietic stem cell repopulation. Other studies implicate its role in leukemia and colon cancer. Downstream targets of the MSI2 homologue MSI1, have shown a strong correlation with cancer related processes through their roles in cell cycle modulation, proliferation, differentiation, and apoptosis. MSI1 has also been shown to modulate WNT and Notch signaling through its interactions with the Notch inhibitor NUMB and the WNT antagonist DKK3.

Invasion, migration, and proliferation assays were carried out in the breast cancer cell lines MDA231 and MCF7 over-expressing MSI2. These cell lines were also transfected with the TOPflash TCF reporter plasmid to investigate the effect of MSI2 over-expression on WNT signaling. To investigate the effects of WNT signaling on MSI2, MDA231 cell lines were treated with the human WNT agonist WNT-3a and MSI2 levels were quantified via western blotting. RNA immunoprecipitation was performed to analyze the downstream targets of MSI2. To investigate the role of MSI2 in breast stem and progenitor cell populations, cell lines were grown as non-adherent mammospheres. MSI2 RNA levels in mammospheres were compared via qRT-PCR. Lastly, the expression of a panel of stem cell markers was compared between cells over-expressing MSI2 and GFP controls via qRT-PCR.

Our results show MSI2 over-expressing MCF7 and MDA231 cells have increased levels of proliferation, migration, and invasion. MSI2 over-expressing cells also up-regulate several putative stem cell markers including Epithelial Cell Adhesion Molecule (EpCAM), CD133, and Aldehyde Dehydrogenase. MSI2 levels are significantly enhanced in mammosphere culture and MSI2 over-expressing cells have demonstrated an increase in mammosphere forming capabilities.

We believe MSI2 plays an important role in breast cancer through its modulation of RNA translation and subsequent regulation of WNT signaling and stem cell biology. MSI2 represents a promising therapeutic target which could be used to block breast cancer invasion and metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2356. doi:10.1158/1538-7445.AM2011-2356
Abstract 2361: The role of thrombomodulin in bladder cancer
Authors: Ding Ann-ann , Su Wen-Pin , Yang Wen-Horng
Published: June 2012

Bladder cancer is one of the most common cancers in the United States and Taiwan. We have discovered that thrombomodulin (TM) is up-regulated in bladder cancer patients and cancer cell lines but not in normal cells. TM is an exclusive endothelial cell surface glycoprotein and an important vascular protective molecule that has anticoagulant, anti-inflammatory and anti-apoptotic properties. TM immunohistochemical analysis of tissue microarray from bladder cancer patients of the United States was performed, and we found that TM staining was more pronounced in high grade and muscle invasive cancers. TM expression by western blot have been examined in 10 bladder cancer tissues from Taiwan in comparison to 3 normal bladder tissues, 80% (8/10) of the cancer group were over-expressed while none showed in normal bladder tissues. In a cell model, we knocked down the TM expression in BFTC905 bladder cancer cells (which was a cell line from invasive TCC) by siRNA. The two stable TM knocked-down cells (C6 and E2) showed a lower proliferation rate than BFTC905 wild type cells and vector control cells (D3). TM knocked-down cells (C6 and E2) also had lower migration and invasion abilities than control cells. Furthermore, TM knocked-down cells have a decreased colony number and size in tumor formation assays. By contrast, SV-HUC-1(immortalized human normal urothelial cell line) enhanced the migration and invasion ability by transient over-expression with TM plasmid. In an animal model, the RT-PCR analysis of the ascites from mice with peritoneal inoculation of MBT-2 cells (mouse bladder tumor cell line), we found that TM expression has higher in the group with tumor formation than the group without. These findings suggest that TM may be involved in the tumor progression of bladder cancer. Thus TM could be a bladder cancer prognosis biomarker and also be a therapeutic target.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2361. doi:10.1158/1538-7445.AM2011-2361
Abstract 2364: Regulation of cofilin activity and tumor cell migration through protein kinase D
Authors: Spratley Samantha J. , Storz Peter
Published: June 2012

During early metastatic events in cancer, reorganization of the actin cytoskeleton must occur to allow for cell invasion and migration. A key enzyme initiating actin remodeling at the leading edge is Cofilin, a small actin binding protein of the ADF/cofilin family with actin severing activities. Cofilin activity is tightly regulated by phosphorylation and dephosphorylation events that are mediated by LIM Kinase (LIMK) and the phosphatase Slingshot (SSH), respectively. Protein kinase D (PKD) is a serine/threonine kinase that has previously been shown to regulate actin driven directed cell migration through phosphorylation and thus inactivation of Slingshot. Here we show that PKD can also regulate cofilin activity via activation of LIMK. We further show that PKD-induced regulation of LIMK and the resulting diminished cofilin activity translates to altered cell motility. Our data suggest that PKD inactivates Cofilin through modulation of both of its regulatory pathways. Further elucidation of the mechanisms by which PKD regulates directed cell migration could enable development of a potential treatment strategy for invasive cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2364. doi:10.1158/1538-7445.AM2011-2364
Abstract 237: MET receptor is a potential new target for cervical carcinoma treatment
Authors: Miekus Katarzyna , Pawlowska Marta , Majka Marcin
Published: June 2012

Introduction: Cervical carcinoma (CC) is the major cause of death among women suffering from neoplastic disease especially in the developing countries. The highest mortality is observed in patients with late diagnosis and metastatic disease. MET protooncogene encodes a tyrosine kinase receptor for hepatocyte growth factor (HGF). MET activation triggers a complex biological program including stimulation of cell proliferation, motility and protection from apoptosis. This program plays an important role during embryonic development and wound healing, but abnormal MET activity contribute to cancer development and progression. Therefore, new therapeutic strategies inhibiting MET signaling have been sought.

Objectives: The aim of this study was to examine the influence of MET inhibition on growth and metastatic ability of cervical carcinoma cell.

Materials and Methods: CC cell line (HTB35) was transduced with MET shRNA and LacZ shRNA expressing viruses and selected with blasticidin. To check genes expression real-time PCR, western blot and flow cytometry were used. Proliferation, cell cycle and apoptosis was studied. Stress fibers organization, adhesion and migration were evaluated using using falloidin staining, adhesion assay and chemotaxis assay respectively. The level of various protein phosphorylation was estimated by western blot.

Results: We showed that downregulation of MET expression in HTB-35 cell line causes: (i) the reduction in cell proliferation resulting from altered cell cycle progression, but not from enhanced apoptosis, (ii) changes in actin cytoskeleton organization, (iii) the increased expression of E-cadherin, (iv) the decrease of mRNA levels in the genes responsible for malignant phenotype (such as c-myc, CXCR4 and Slug), and (v) the increased expression of genes involved in EMT and metastasis (including Snail, Slug, Twist, MMP-2, MMP-9 and HIF-1α). Performed in vitro studies have not shown any significant effect of MET inhibition on adhesive ability of HTB-35 cervical carcinoma cell line. However, the decreased migration of MET negative cells was observed.

Summary: Our results strongly, indicate that MET receptor plays an important role in stimulating growth and metastasis of HTB-35 cervical carcinoma cell line. Thus, inhibition of HGF-MET receptor axis could be used in the future as the new therapeutic strategy to block cervical carcinoma development and progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 237. doi:10.1158/1538-7445.AM2011-237
Abstract 2370: Single cell analysis using a microscale migration slide
Authors: Zhou Yi , Moore Cristina , Chu Vi
Published: June 2012

Cell migration occurs in a variety of biological processes including wound healing, embryonic development, and immune responses. Existing chemotaxis assays used to assess cell migration employ the Boyden chamber which lack real-time imaging capability and does not maintain well defined biomolecular gradients. To overcome these limitations, a cell migration assay was developed which enabled real-time monitoring of single cells on a slide-like platform that maintained a stable linear gradient. Fluorescence measurements with albumin (MW=66kDa) conjugated with FITC demonstrated that the concentration profile was linear over the 1 mm observation area. This linear concentration gradient was established after 1 hour incubation and was stable for over 24 hours.

The microscale migration slide was used to assess the effects of serum concentrations (0, 2, 5, 10, and 15%) on the migration propensity of four different cell lines that were grown on collagen. The cell lines included cancer cell lines with known invasive properties (fibrosarcoma cancer cells HT1080 and breast cancer cells MDA-MB-231), along with HUVECs which are angiogenic cells involved in tumor metastasis and a control cell line, NIH 3T3 fibroblast cells. Cells were incubated in the experimental conditions overnight (12 hours) with images captured every 15 minutes. Migration of single cells was monitored and imaged in real-time and the percentage of cells that exhibited positive migration towards the chemotactic source was calculated. Interestingly, we observed that the highest percentage of migratory HUVEC cells was obtained using 2% serum whereas the other three cell lines exhibited increased migration with increasing serum concentration. Under similar culture conditions of either 10 or 15% serum, MDA-MB-231 possessed the highest migration index, followed by NIH 3T3 fibroblasts and HT1080 cells whereas HUVEC cells did not migrate at all under these high serum conditions. Extensive testing over a range of conditions with different users indicated remarkable consistency in the data generated and suggests that the migration slide is a useful tool to monitor the effects of chemoattractants on adherent cell lines. Overall the results provide a reference point from which to build upon future studies aimed at comparing the effects of signaling molecules and growth factors on the migration propensities of cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2370. doi:10.1158/1538-7445.AM2011-2370
Abstract 2388: Targeted expression of BAG3 to the murine mammary gland (MMTV-hBAG3) reveals a role in mammary tumorigenesis
Authors: Casagrande Giovanna , Yu Minshu , Rodriguez-Canales Jaime , Kohn Elise C. , Virador Victoria M.
Published: June 2012

We have previously shown that BAG3 is a pro-survival protein in human cancer lines subject to stress. BAG3 overexpression also mediates a decrease in migration and adhesion to matrix in MDA-435 breast cancer cells; this is reversed in a BAG3 mutant with a deleted proline-rich domain (PXXP). Additionally, BAG3 induces the secreted molecule CCN-1 to regulate in vitro invasion of HeLa cells. This led us to hypothesize that MMTV-driven human BAG3 would contribute to mammary tumorigenesis. This was examined through generation and analysis of a transgenic mouse model of human BAG3 ectopically expressed under the MMTV promoter in FVB/N mice. Five of 14 confirmed founder lines were selected for characterization to rule out integration effects. Each line was shown to express hBAG3 in mammary ductal epithelium, a typical feature of the MMTV-driven transgenes. We next applied chemical carcinogenesis with DMBA (7,12-dimethylbenzanthracene) to eight-week old virgin transgenic females and age-matched FVB/N wild-type females, by weekly orogastric gavage (1 mg DMBA) for 6 weeks. At study end (18 weeks post DMBA-treatment), 5/19 (26%) transgenic animals had pathologically proven atypical hyperplasia [1], and malignant tumors [adenocarcinoma, 1; myoepithelial tumor, 2; adenosquamous carcinoma, 1]. The age-matched non-transgenic controls did not develop premalignant or malignant breast lesions [0/12]. Luminal cell expression of hBAG3 was present in the atypical hyperplasia, but was not observed in highly proliferative, less differentiated regions of tumors where ductal structures are absent. Since the tumors we obtained were both of the myoepithelial and the epithelial lineages, we next investigated whether forced expression of hBAG3 in the ductal epithelium may dysregulate mammary gland development. We collected mammary glands at 5, 7, 10, and 13 weeks of age from transgenic and wild-type animals (n=4 each per time point) for two selected MMTV-hBAG3 transgenic lines. The terminal end buds, epithelial elongation, and glandular differentiation are currently being analyzed. Our preliminary data indicate that the mammary gland of MMTV-hBAG3 mice have a trend to increased epithelial elongation and more secondary acinar structures in the later pubertal stages. A complete analysis of the study cohort will be discussed. The effects of multiple pregnancies and post-lactation involution are under study. Together our findings point to a putative role of BAG3 as contributor to mammary remodeling during puberty and a promoting effect during chemical carcinogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2388. doi:10.1158/1538-7445.AM2011-2388
Abstract 2390: Growth analysis of M-406 mammary adenocarcinoma in F1 and F2 generations of a cross between resistant and susceptible mice lines
Authors: Cáceres Juan M. , Pagura Lucas , Zacarías-Fluck Mariano F. , Scharovsky O Graciela , Di Masso Ricardo J. , Rico María J. , Rozados Viviana R.
Published: June 2012

Breast cancer is a complex, multifactor disease that affects about 10% of women in industrialized countries. Inbred mice models provide an interesting tool for identifying genes that control susceptibility to breast cancer. CBi- mice was artificially selected for body conformation from CBi. The mammary adenocarcinoma M-406 developed spontaneously in CBi female mouse and is maintained in vivo in mice of the same line. When CBi- mice are s.c. challenged with M-406 they show 100% take and 100% regression (resistance). In contrast, the tumor grows exponentially (susceptibility) in CBi mice. The aim of this study was to characterize the tumor behavior in both F1 reciprocal hybrids [(CBi × CBi-) and (CBi- × CBi) n=48], and in the four F2 combinations between them: F2A [(CBi- × CBi) × (CBi- × CBi)]; F2B [(CBi- × CBi) × (CBi × CBi-)]; F2C [(CBi × CBi-) X (CBi- × CBi)] and F2D [(CBi × CBi-) X (CBi × CBi-)] n=185. Adult female and male mice where challenged s.c. with M-406 by trocar, three tumor fragments (∼8 × 105cells). Tumors were measured three times/week and tumor volumes were calculated as follows: V = 0.4 (ab2), where V = volume (mm3), a = largest diameter (mm) and b = smallest diameter (mm). Tumor volume vs time data were fit to an exponential growth model Vt: S*e[k*t]. M-406 grew exponentially in 45/48 F1 mice. Non statistical differences were found in tumor growth rates between sexes on the reciprocal F1s. Significant differences in tumor volume doubling time (TvDT = 0.69/k) were observed between sexes in two of the four F2 combinations: F2A [females: 5.34 ± 0.44; males: 4.27 ± 0.26; t=2.154, P=0.038] and F2D [females: 4.04 ± 0.14; males: 4.85 ± 0.38; t=1.804, P=0.041]. When comparing females TvDT, F2D had a faster growth rate than F2A (P=0.0097), while non statistical differences were evident in males (P = 0.102). It is concluded that susceptibility to M-406 in CBi mice is inherited as a dominant trait. Genes involved in susceptibility inheritance would be autosomic, as the tumor behaviour in both F1 reciprocal hybrids is similar. The analysis of F2 alternative combinations allowed the identification of maternal effect as well as sexual dimorphism influencing tumor growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2390. doi:10.1158/1538-7445.AM2011-2390
Abstract 24: A genetic model of metastatic evolution: Driver and passenger mutations affect metastatic fitness
Authors: McFarland Christopher D. , Scott Jacob G. , Basanta David , Anderson Alexander R. A. , Mirny Leonid A.
Published: June 2012

Background: The metastatic process requires that cells accomplish two feats: 1) migrate from the primary tumor into the blood stream and arrest in foreign stroma, and 2) grow from a colony of only a few cells into a metastasis of macroscopic size. This second step is highly inefficient for reasons not well understood. Our previous research in tumorigenesis found that many mildly deleterious passenger mutations accumulate in cancer and can influence neoplastic progression, yet the effect of these mutations in metastatic evolution has not been studied.

Methods: We developed a model of somatic evolution of neoplastic progressions and metastatic growth. Using a modified Gillespie algorithm, individual cells in our model stochastically divide and die, occasionally acquiring advantageous driver mutations or mildly deleterious (for cancer) passenger mutations. The rate of cell division and death depend on the collective effect of accumulated passenger and driver mutations. Evolutionary pressures lead to clonal expansion and occasionally carcinogenesis. In our in silico experiments, aliquots of cells were taken from successful primary tumors and “injected” into a new micro-environment with new stromal interactions, where they were assayed for metastatic success. This allowed us to track the entire evolutionary history of metastatic cells and genomic determinants of their success.

Results: Accumulation of passengers during tumorigenesis as well as metastatic progression frequently prevents initial colonies and micrometastases from developing into metastatic tumors. The population bottleneck experienced during colony formation accelerates the accumulation of deleterious passenger mutations in cancerous populations. We find that in the model, metastatic efficiency depends heavily on primary tumor size, age, and genetic diversity in a manner consistent with clinical observations. A favorable stromal environment was also critical for metastatic success. Surprisingly, we found that as the total number of mutations in neoplastic cells increased, metastatic efficiency, on average, decreased as many of these additional mutations were deleterious passengers.

Conclusions: Accumulation of deleterious passenger mutations helps explain metastatic inefficiency after extravasation, as well as many other features of metastasis. Our model makes several novel predictions testable in vivo_most notably that increased mutational load may prevent or impede metastasis. Cancer treatments exploiting the deleterious effects of passenger mutations may prevent metastatic disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 24. doi:10.1158/1538-7445.AM2011-24
Abstract 2406: Molecular characterization of ciclooxigenase-2 in canine proliferative inflammatory atrophy of the prostate
Authors: Amorim Renee L. , Rodrigues Marcela , Rosa Luciana , Drigo Sandra , Cirilo Priscila , Rogatto Silvia
Published: June 2012

Due to the importance and incidence of prostatic carcinoma in man, some animals are being used as a model in the investigation of the pathogenesis and development of this lesion. Similarly to men, the dog presents a spontaneous development of prostate cancer, which is locally invasive and can develop bone metastasis. Proliferative inflammatory atrophy (PIA) is considered a pre-neoplastic lesion in men. Overexpression of COX-2 has been associated with PIA, PIN and prostatic carcinoma playing a role in angiogenesis, apoptosis inhibition, increasing of invasive capacity, metastasis and inflammation in different prostatic affections. The aim of this study was to assess the protein and gene expression of COX-2 in laser capture microdissected stroma and epithelial cells of PIA foci in canine prostate by immunohistochemistry (IHC) and qRT-PCR. For IHC, 38 PIA lesions arranged in a tissue microarray were evaluated. Frozen fragments from 17 PIA and 3 normal prostatic tissues were used for COX2 gene expression. Our results showed increased COX2 gene expression in stromal cells compared to epithelial cells. High level of COX-2 protein expression was observed in 87% of PIA lesions, despite epithelial or stromal compartment. The In the present study, the stroma presented an important role, since the COX-2 gene expression in canine PIA lesions was increased in this cell compartment. To the best of our knowledge, this is the first report of COX2 gene expression in canine PIA. Since high levels of COX-2 have been associated to the risk of developing of prostate cancer in man, dog can be used as a natural model for this disease. The stroma can act as paracrine way in the epithelial cells, stimulating epithelial proliferation, reducing the apoptosis of this compartment and promoting angiogenesis in the microenvironment, factor that contribute to prostatic carcinogenesis and that the protein expression is increased in this lesion.

Sponcer: Fapesp (Fundação de Apoio a Pesqusia do Estado de São Paulo)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2406. doi:10.1158/1538-7445.AM2011-2406
Abstract 2417: A new mouse model for epithelial ear neoplasms based upon expression of mutant EGFRL858R/T790M
Authors: Kawabata Shigeru , Hollander M. Christine , Munasinghe Jeeva P. , Mercado Jose , Brinster Lauren R. , Butman John A. , Lonser Russell R. , Regales Lucia , Pao William , Janne Pasi A. , Wong Kwok-Kin , Dennis Phillip A.
Published: June 2012

Epithelial neoplasms of the ear are rare and difficult to detect before the development of audiovestibular or facial nerve dysfunction. Complete resection is often difficult because of location and the late stage at diagnosis, so effective chemotherapy is required. No preclinical models exist for these tumors. During the generation of mice using CC10 or surfactant protein C (SP-C) promoters to drive expression of mutant EGFR (EGFRL858R/T790M) in lung tissues, we observed that no SP-C mice developed lung tumors, but approximately 30% of them developed head tilt and a circular gait by 25 wk. MRI scans showed bilateral ear tumors located in the tympanic cavity that partially invaded into temporal bone, which suggested the presence of middle ear tumors or endolymphatic sac tumors (ELSTs). Histology of the ear tumors showed well-differentiated adenocarcinomas with multiple papillary and microcystic structures. Immunohistochemical (IHC) analysis showed that the mutant EGFR was active and selectively expressed in the tumors, and that downstream components of EGFR signaling such as Akt, mTOR and ERK were activated. SP-C expression was also detected, suggesting that surfactant proteins are expressed in ear epithelium at specific times during tumor development. To determine if these tumors were addicted to EGFR signaling, EGFR-based therapies were tested in symptomatic mice. Consistent with the T790M mutation in EGFR conferring resistance to reversible EGFR tyrosine kinase inhibitors (TKI), erlotinib was ineffective. In contrast, the combination of an irreversible EGFR TKI (BIBW-2992) with an antibody against EGFR (cetuximab), or a chemically optimized analogue of BIBW-2992 (WZ4002) used alone, was highly effective. All mice treated with the combination or WZ4002 alone had normalization of gait and complete responses by MRI, suggesting that these tumors are dependent upon mutant EGFR. To assess the possible clinical relevance of this mouse model, 2 human middle ear adenocarcinomas and a human ELST were stained for active EGFR using IHC. 3/3 displayed increased EGFR activation. These studies describe a new mouse model for human epithelial ear tumors based on EGFR activation, and raise the possibility that EGFR-based therapies might have clinical efficacy in these rare neoplasms.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2417. doi:10.1158/1538-7445.AM2011-2417
Abstract 243: Functional dissection of hoxD genes in regulation of neuroblastoma cell proliferation and differentiation
Authors: Zha Yunhong , Ding Emily , Mao Ling , Ding Han-fei
Published: June 2012

Neuroblastoma is a common malignant tumor in children, arising in the sympathetic nervous system. The tumor cell differentiation state influences clinical outcome, and induction of differentiation has been considered as a promising therapeutic strategy for neuroblastoma. Retinoid acid (RA) is an effective inducer of neuroblastoma cell differentiation and has been used in clinic for treatment of high-risk neuroblastomas. However, resistance to RA presents a major obstacle to successful RA-based therapy. Therefore, identification of target genes that mediate the anti-neoplastic activity of RA is of paramout importance, as targeting these genes offers opportunities to bypass resistance to RA. A variety of experimental evidence has linked RA action to the activation of HOX genes, which function as master regulators of development and cell fate specification. Particularly, it has been shown that RA induces several HOXD genes in neuroblastoma cells including HOXD1, D8 and D9. Our microarray analysis confirmed the predominant upregulation of HOXD genes in the human neuroblastoma cell line BE(2)-C cells following RA treatment. To investigate the functional significance of the RA-induced upregulation of HOXD genes, we conducted a comprehensive functional analysis of HOXD genes in regulation of neuroblastoma cell proliferation and differentiation. The HOXD cluster contains 9 genes (HOXD1, D3, D4, and D9-13) that are positioned from 3’ to 5’ within the cluster, with HOXD1 at the 3’ end and HOXD13 the 5’ end. We established BE(2)-C-derived cell lines with inducible expression of individual HOXD genes. Induction of HOXD1, D12 or D13 had no significant effect on BE(2)-C cell growth and differentiation. By contrast, individual induction of other HOXD genes caused G1 arrest, which was associated with downregulation of cyclins and CDKs required for cell cycle progression through the S, G2 and M phases. Ectopic expression of HOXD3, D4, D8, D9, D10 or D11 also induced morphologic changes characteristic of neuronal differentiation. Interestingly, HOXD8 and D9 appeared to be most potent in the induction of G1 arrest and differentiation. These findings suggest that HOXD genes positioned in the middle of the cluster have higher differentiation-inducing activity compared to those positioned more 5’ and 3’ in the cluster. Thus, pharmacological activation of HOXD8 and HOXD9 may represent a therapeutic strategy for RA-resistant neuroblastomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 243. doi:10.1158/1538-7445.AM2011-243
Abstract 2441: CHI3L1 in allergic pulmonary inflammation promotes breast cancer metastasis by upregulation of MMP-9
Authors: Libreros Stephania , Garcia-Areas Ramon , Iragavarapu-Charyulu Vijaya L.
Published: June 2012

Disseminated metastasis is one of the main causes of malignancy related deaths in breast cancer patients. Murine mammary tumors cells metastasize to the lung 5 weeks post tumor-implantation. However, in the presence of asthma, metastasis is seen at an earlier time point. In this study, we investigated the molecule(s) contributing to early metastasis. One of the common molecules among asthma and breast cancer is a glycoprotein known as chitinase-3-like-1 (CHI3L1) which induces TH2 proinflammatory cytokines and matrix degrading enzymes. The expression of CHI3L1 is elevated in the lungs and serum of asthmatic patients and correlates with disease severity. Interestingly, this glycoprotein is also elevated in plasma of patients with metastatic breast cancer. Using murine mammary cancer models, we show the link between increased metastasis and asthma. The pathways that tumor cells use for metastasis have significant similarities with the mechanisms mediating allergen-induced leukocyte recruitment. Thus, leukocyte recruitment to the site of inflammation and tumor cell metastasis involves extracellular matrix degradation. Matrix metalloproteinase-9 (MMP-9) is one such enzyme that can degrade all aspects of the extracellular matrix. Our previous studies have shown increased levels of MMP-9 in mammary tumor-bearing mice. MMP-9 has been reported to be upregulated in the presence of CHI3L1. In this report, we show that the levels of CHI3L1 are higher in asthmatic tumor bearers compared to either tumor controls or asthma controls. We hypothesize that the elevated CHI3L1 expression causes increased MMP-9 production leading to higher metastasis. To answer this hypothesis, a mouse model of allergic pulmonary inflammation and two types of well-established metastatic mammary tumor models were used. Analysis of serum, bronchioalveolar lavage (BAL), lung and spleen by RT-PCR and ELISA revealed higher expression of CHI3L1 in asthmatic tumor bearers. In summary, these studies implicate a relationship between allergic pulmonary inflammation and increased breast cancer metastasis to the lung. As CHI3L1 is involved in both of these diseases, this molecule may be a novel target for suppressing/preventing metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2441. doi:10.1158/1538-7445.AM2011-2441
Abstract 245: KIF1Bβ interacts with YME1L1, resulting in apoptosis accompanied by the alteration of mitochondrial morphology
Authors: Ando Koji , Tokochi Tomoki , Mukai Akira , Maehara Yoshihiko , Nakagawara Akira
Published: June 2012

We previously reported that KIF1Bβ is a tumor suppressor gene mapped to chromosome 1p36.2, and is related to apoptosis (JBC, 2008). It is known that cell survival and apoptosis have a close relationship with mitochondrial morphology, such as fusion and fragmentation. Cell survival and resistance to pro-apoptotic stimuli is related to mitochondrial fusion. On the other hand, apoptosis is frequently associated with mitochondrial fragmentation. Considering these facts, we hypothesized that KIF1Bβ might play an important role in the morphological change in mitochondria.

The mitochondrial morphology was examined by indirect immunofluoresence using MitoTracker Red® and anti-cytochrome c antibody. To quantify the degree of mitochondrial fusion, we used JC-1 probe. Yeast two hybrid screening was performed to identify the interacting proteins of KIF1Bβ.

Mitochondrial fusion was facilitated in both NB1 (neuroblastoma cell line which KIF1Bβ is homozygously deleted) and HeLa in which KIF1Bβ was knocked down. This result suggests that KIF1Bβ takes part in the process of morphological change in mitochondria. Those cells were resistant to doxorubicin treatment, demonstrating that apoptosis was prevented accompanied by mitochondrial fusion. Overexpression of KIF1Bβ induced mitochondrial fragmentation, enhanced cytochrome c release into the cytoplasm, and resulted in apoptosis. These results are consistent with the properties of KIF1Bβ as a tumor suppressor described previously. On the other hand, overexpression of KIF1Bβ lacking the death inducing region neither induced mitochondrial fragmentation nor apoptosis, indicating that the death inducing region is essential for the alteration of mitochondrial morphology and induction of apoptosis. Utilizing yeast two hybrid screening, we found several proteins physically interacting with KIF1Bβ. We identified YME1L1, the mitochondrial metalloprotease, which is known to cleave mitochondrial fusion protein OPA1. Overexpression of YME1L1 cleaved OPA1 in the presence of KIF1Bβ. However, OPA1 cleavage was partially inhibited in the absence of KIF1Bβ, suggesting that KIF1Bβ enhances the activity of YME1L1 as a protease to cleave OPA1. Interestingly, high expression of YME1L1 was associated with better survival of neuroblastoma patients (n=101; p=0.046), while high expression of OPA1 was correlated with poor outcome (n=101; p=0.04).

These results suggest that the tumor suppressor KIF1Bβ takes part in the alteration of mitochondrial morphology collaborating with an interacting partner YME1L1.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 245. doi:10.1158/1538-7445.AM2011-245
Abstract 2454: Pancreatic cancer stem cells (CSCs) express MUC1 and MUC1-expressing pancreatic cancers encompass higher levels of CSCs
Authors: Curry Jennifer M. , Thompson Kyle J. , Rao Shanti , Ahrens William A. , McKillop Iain H. , Sindram David , Iannitti David A. , Martinie John B. , Mukherjee Pinku
Published: June 2012

Introduction: Pancreatic ductal adenocarcinoma (PDA) has the worst prognosis of all cancers and is the fourth leading cause of cancer-related deaths in the United States. Cancer stem cells (CSCs), which are a small population of tumor cells that have the ability to self-renew and differentiate into the diverse cells that comprise the tumor, have been identified in pancreatic cancer patients as lineage-negative, CD133 (prominin-1)-expressing cells. In pancreatic cancer, MUC1 is a protein expressed with a high frequency and its expression correlates with high metastasis and poor prognosis. Recent reports from our lab have demonstrated the ability of MUC1 to drive epithelial-to-mesenchymal transition thus producing a cell with ‘stem-like’ properties. We therefore investigated the correlation between MUC1 expression and CSCs. Methods: Primary tumor tissues surgically resected from patients diagnosed with pancreatic adenocarcinoma (n=8) and human pancreatic cancer cell lines were assessed for CD133 and MUC1 expression by flow cytometry. Results: In pancreatic cancer patients, tumor MUC1 levels correlated with a higher percentage of CD133-expressing cells (R=0.76). Similarly, in MiaPaCa2, Capan-1, BxPC3 WT, BxPC3 MUC1, Su86.86 WT and Su86.86 MUC1 cells lines, MUC1 levels correlated with increased CD133 expression (R=0.83). The majority of CD133+ cells isolated from primary human pancreatic tumors (82.6 ± 12.7%) expressed MUC1. Similar results were obtained using cell lines, as 83.22 ± 30.9% of CD133-expressing cells were MUC1-positive. Conclusion: MUC1 is present on the majority of CSCs and its expression correlates with higher levels of cancer stem cells. This study demonstrates yet another oncogenic role of MUC1 in pancreatic cancer progression and suggests MUC1 as a novel marker for CSCs.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2454. doi:10.1158/1538-7445.AM2011-2454
Abstract 2476: Acetylated Survivin functions in DNA damage repair
Authors: Cooper Zachary A. , Riolo Matthew , Holloway Michael P. , Altura Rachel A.
Published: June 2012

Survivin is an oncogenic protein that participates in cell division and inhibits apoptosis in cancer cells. These functions are dependent on its interaction with partner proteins within distinct subcellular compartments. Survivin is highly expressed in malignant tumors yet minimally expressed in normal tissues, making it an attractive target for molecular therapy. Inhibition of Survivin function enhances tumor cell sensitivity to radiation, the mechanism of which has been reported to result from a disruption of DNA-double-strand break repair processes. Recently, we demonstrated that Survivin is post-translationally modified by acetylation at numerous lysine residues, one of which is responsible for its subcellular transport. Acetylation at Lys-129 facilitates Survivin homodimerization which prevents its binding to the CRM1 nuclear export protein, inhibiting its translocation from the nucleus to the cytoplasm. The object of this study was to determine the potential relationship between Survivin acetylation and its function in nuclear repair processes. To elucidate the association between Survivin acetylation, nuclear localization and DNA double-strand break repair we subjected HeLa cells to 4Gy γXRT and examined Survivin acetylation at serial time points (20, 40, 60 min) after irradiation. An immediate deacetylation of Survivin was observed at 20 minutes, followed by an increase in both global acetylation and acetylation at Lys-129 at 40 minutes. The increase in Lys-129 acetylation correlated with its nuclear accumulation, as demonstrated by indirect immunofluorescence staining and immunoblotting after subcellular fractionation. Acetylated Survivin was found in endogenous immunoprecipitants with the DNA damage repair protein, Ku70, following γ-irradiation. Indirect immunoflourescence revealed co-localization of Ku70 and acetylated Survivin within the nucleus at similar time points after irradiation suggesting that Survivin acetylation may be required for Ku70 complex formation within the nucleus. These results implicate acetylated Survivin as a regulator of DNA damage repair through the Ku70 DNA repair pathway and further our understanding of Survivin function within the nuclear compartment of cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2476. doi:10.1158/1538-7445.AM2011-2476