bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3261: GRP78 regulates clusterin stability, retrotranslocation, and mitochondrial localization under ER stress in prostate cancer
Authors: Li Na , Zoubeidi Amina , Beraldi Eliana , Gleave Martin E 
Published: August 2012

Abstract
Expression of clusterin (CLU) closely correlates with regulation of apoptosis in cancer. While endoplasmic reticulum (ER) stress-induced up-regulation and retro-translocation of cytoplasmic CLU (cCLU)(also termed as presecretory and secreted forms, psCLU and sCLU) has been linked to its anti-apoptotic properties, the mechanism mediating these processes remain undefined. Here, we show that GRP78 (Bip) associates with cCLU to facilitate its retro-translocation and redistribution to the mitochondria in human prostate cancer cells. We found that ER stress inducers, such as TG or MG132, increased expression levels of both cCLU and GRP78 proteins and led to the association of these two proteins. Furthermore, GRP78 overexpression resulted in increased cCLU but reduced sCLU levels secreted into the culture media. GRP78 stabilized cCLU protein and its hypo-glycosylated forms, in particular after paclitaxel treatment. Moreover, subcellular fractionation studies indicated GRP78 increased stress-induced cCLU retro-translocation from the ER with redistribution to the mitochondria. Live cell imaging revealed that a CLUGFP localized directly to highly mobile mitochondria. Co-expression of both GRP78 and CLU in LNCaP cells led to increased CLUGFP -mitochondria co-localization and reduced apoptosis by cooperatively stabilizing mitochondrial membrane integrity. Transient depletion of GRP78 reduced cCLU protein but not mRNA transcript levels, accompanied by cell cycle arrest at G0/G1 with down-regulation of both phospho-Rb and cyclin D1, and synergistic increases in paclitaxel-induced cell apoptosis. Taken together, our findings reveal a novel role of GRP78 in the regulation of CLU retro-translocation and its redistribution to the mitochondria, suggesting that GRP78 and CLU cooperatively promote survival during ER stress in prostate cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3261. doi:1538-7445.AM2012-3261
Abstract 3264: Downregulation of intersectin-1s in human lung cancer may contribute to tumorigenesis
Authors: Predescu Dan , Zhang Jin , Patel Monal , Bardita Cristina , Predescu Sanda 
Published: August 2012

Abstract
The expression of intersectin-1s (ITSN-1s), a granzyme B (GrB) substrate and one of the trisomic genes that may provide protection against most cancers in Down's syndrome, is significantly downregulated in human lung cancer (LC) cells and tissue. To date, there is no information on the cellular and molecular factors that modulate ITSN-1s expression or whether downregulation of ITSN-1s can contribute to LC. Thus, we hypothesize that loss of full-length ITSN-1s due to proteolytic cleavage by GrB during an inflammatory response and the interplay between the signaling events triggered by the two cleavage products, are part of a complex common molecular mechanism involved in the oncogenic transformation of cells. To address this hypothesis, we used cultured LC cells and tissue, as well as a range of biochemical (Western blot, ELISA, BrdU incorporation), molecular biology (plasmid DNA transfection, qPCR), morphological (immuno-histochemistry, fluorescence microscopy) approaches, soft agar colony formation and xenograft tumor formation in nude mice to evaluate whether downregulation of ITSN-1s can contribute to lung carcinogenesis. Western blot and immuno-histochemistry of LC cells and lung tissue lysates showed significant decrease in ITSN-1s protein levels, while qPCR revealed only slight decrease, if any, in ITSN-1s mRNA. In vitro studies showed that GrB cleaves ITSN-1s and generates a cleavage fragment (GrB-EHITSN) with high proliferative potential. Lung epithelial cells expressing the GrB-EHITSN show 40% increase in BrdU incorporation, selective activation of p38MAPK and downstream Elk-1 transcription factor. These findings are consistent with a p38MAPK and Elk-1 pro-proliferative signaling pathway regulated by ITSN-1s. Activation of p38 and Elk-1 was also detected by western blot using phospho-specific antibodies, in all LC cells investigated. Expression of the two GrB cleavage fragments of ITSN-1s, individually or together, in human lung epithelial cells exerts a modulatory effect on the balance p38/Erk1/2MAPK activation. Functional studies showed that ectopic expression of ITSN-1s in the LC cells restored the p38/Erk1/2MAPK balance, significantly reduced their proliferation rate as well as the formation of colonies in soft agar and xenograft tumors in nude mice. Altogether, our results suggest that downregulation of ITSN-1s can promote the oncogenic transformation of cells.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3264. doi:1538-7445.AM2012-3264
Abstract 3265: PMA induces malignant characteristics in MCF-7 breast cancer cells via MAPK-pathway
Authors: Hautala Laura C. , Koistinen Riitta , Koistinen Hannu 
Published: August 2012

Abstract
Phorbol 12-myristate 13-acetate (PMA) is a well known tumor promoter, the effect of which is mainly mediated by protein kinase C (PKC). The effect of PMA varies depending on the cell line and growth conditions. In MCF-7 breast cancer cells PMA has been found to induce growth arrest and resistance to apoptosis. Here, we studied the effects of PMA on differentiation and malignant characteristics of MCF-7 cells. The cells were grown on Matrigel besement membrane preparation or on plastic/glass surface with and without PMA, rottlerin or bryostatin. The cell proliferation assay was carried out using the cells grown on 96-well plates. Wound healing test, on glass surface, was used to detect the differences in cell migration. Activation of MAPK-pathway proteins was studied with Phospho-kinase array. When grown on Matrigel, the addition of PMA to cell culture medium increased the motility of the cells and induced dramatic morphological changes, leading to the formation of web-like multicellular structures. These changes were not observed when the cells were grown on plastic or glass surface. However, PMA caused a two-fold increase in the migration of the cells on glass surface. Furthermore, the PMA treated cells had lower proliferation rate than the untreated cells. Preincubation of the cells with PKC-inhibitors rottlerin or bryostatin abolished the effects of PMA, suggesting that these effects are mediated by PKC. PMA also induced several changes in the phosphorylation of different proteins involved in MAPK-pathway. Our results confirm the previous findings showing that PMA induces malignant characteristics in MCF-7 breast cancer cells. Morphological differentiation was only seen when the cells were grown on basement membrane preparation, recapitulating the physiological growth environment of the cells better than the artificial plastic surface. Furthermore, our results indicate that the effects of PMA on MCF-7 cells are mediated by MAPK-pathway.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3265. doi:1538-7445.AM2012-3265
Abstract 3269: Basal VEGF protein expression in skeletal muscle, adipose tissue and mammary tumors of PyMT mice
Authors: Vona-Davis Linda , Audet Gerald , McLaughlin Sarah , Olfert Mark 
Published: August 2012

Abstract
Obesity is associated with the overall risk of certain cancers, including breast cancer. The prevailing view is that metabolic alterations associated with obesity result in the production of excess vascular endothelial growth factor (VEGF) thereby establishing a local and systemic environment that favors the development and progression of breast cancer. We need to understand how VEGF from adipose is integrated with mammary tumor growth and vascular activity to modulate tumor progression. The MMTV-PyMT mouse is a clinically-relevant model for studying breast cancer progression as mammary tumors grow spontaneously over time; however, little is known about the distribution of VEGF between various organs. We sought to compare the basal VEGF protein expression in skeletal muscle, adipose tissue and mammary tumors from MMTV-PyMT mice and compared these levels against background nontransgenic FVB/N mice. Normal expression of VEGF protein was 69.4 ± 2.1 pg/mL in skeletal muscle and 59.0 ± 7.3 pg/mL in adipose tissue of wild-type (FVB background) control mice. The average expression level of VEGF protein in mammary tumors of MMTV-PyMT mice was 61.8 ± 31.3 pg/mL. Adipose tissue of MMTV-PyMT exhibited 324% greater (191.2 ± 37.6 pg/mL, p&lt;0.01) VEGF protein expression compared to adipose tissue of control FVB mice, and 309% greater (p&lt;0.05) VEGF protein compared to tumor VEGF expression. These data imply that stromal expression of VEGF may be a critical determinant regulating the angiogenic phenotype in breast cancer, and further suggests that cross-talk between the stromal environment and tumor is likely to be essential in promoting tumor growth and progression. NIH 5T32-HL090610, AHA 10BGIA3630002, ACS 116837IRG0906101
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3269. doi:1538-7445.AM2012-3269
Abstract 3270: The effect of soy exposure in different ages on the mammary tumor development of the MMTV-ErbB-2 transgenic mice
Authors: Feng Xiaoshan , Hou Jianfeng , Gao Shegan 
Published: August 2012

Abstract
Objective:To investigate the effect of different soy exposure timings on the mammary tumor development using MMTV-erbB-2 transgenic mice. Furthermore, the underlying mechanisms were also examined. Methods:120 MMTV-erbB-2 transgenic female mice were randomly divided into four groups:control group, early exposure group, late exposure group and lifelong exposure group. The control group mice were treated with soy free diet from week 3 to week 60, the experiment groups mice were treated with soy diet between week 3 to week 12, week 20 to week 60, week 3 to week 60, respectively. Then, the body weight change of each mice, the latency period, and the growth speed of mammary tumor were recorded. Furthermore, the mammary gland morphology was examined by whole mount at 5 weeks. In order to understand the underlying mechanisms of the mammary tumor development, the signal pathways of c-Fos, erbB2 and p-ERK were also examined in breast cancer tissue using immunohistochemistry. Results:We found that the weight in the four groups had no significant difference. Compared with the control group, the TEB numbers and the growth speed of tumors in the early exposure group and the lifelong exposure group were significantly decreased and the tumor latency periods were significantly prolonged (P&lt;0.05). Furthermore, immunohistochemistry showed that the mice in the early exposure group and the lifelong exposure group displayed decreased expression of c-Fos and p-ERK (P&lt;0.05). The expression of erbB-2 protein among the different groups was not statistically significant (P&gt;0.05). Conclusion: Early or lifelong soy exposure affects the morphological development of mammary gland of MMTV-erbB-2 transgenic mice. Soy may display a preventive effect on the initiation of breast cancer. These obersveration may serve as a guide for soy use in the early prevention of the breast cancer. The c-Fos and and p-ERK may be involved in the development of mammary tumor in the MMTV-erbB-2 transgenic mice. The underlying mechanism warrants further studies.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3270. doi:1538-7445.AM2012-3270
Abstract 3271: Targeting fibroblast growth factor receptor 4 decreases prostate cancer growth and metastasis
Authors: Feng Shu , Yu Wendong , Cai Yi , Wang Jianghua , Ittmann Michael M. 
Published: August 2012

Abstract
FGFR4 up-regulation occurs in advanced prostate cancers. Multiple FGFs have been found upregulated in prostate cancers, including FGF1, FGF2, FGF6, FGF8 and FGF17, all of which can activate FGFR4. Furthermore, a SNP changing the sense codon 388 from glycine to arginine in the transmembrane domain of the human FGFR4 gene has been linked to prostate cancer initiation, progression and poor prognosis. In this study, we determined whether downregulation of FGFR4 expression using small hairpin RNA (shRNA) against FGFR4 inhibits prostate cancer progression in vitro and in vivo. We used two well-characterized prostate adenocarcinoma cell lines, PC3 and LNCaP to derive prostate cancer cells stably expressing a shRNA targeting FGFR4 (PC3-sh-FGFR4 and LNCaP-sh-FGFR4) by using lentivirus vector infection. Real-time quantitative RT-PCR shows that FGFR4 mRNA expression was decreased by 87% (PC3-sh-FGFR4) and 78% (LNCaP-sh-FGFR4) compared to scramble control shRNA cells PC3-shV and LNCaP-shV, respectively. In vitro analysis showed that the targeted knockdown of FGFR4 gene in PC3 and LNCaP cells with sh-FGFR4 resulted in significantly inhibited cell proliferation and invasion. The reduction of Akt phosphorylation on both Ser473 and Thr-308 sites detected by western blot suggests that FGFR4 signaling is implicated in Akt activation. The upregulated expression of Caspase 8 and activated Caspase 3 in FGFR4 knockdown cells measured with immunoblot indicates that FGFR4 signaling inhibits prostate cancer cell apoptosis. The prostate cancer orthotopic xenograft models established with PC3-sh-FGFR4 and scramble shRNA control PC3 cells reveal that FGFR-4 knockdown significantly inhibited tumorigenicity in this model (P=0.01). In mice with pathologically confirmed primary tumors, primary tumor weight was decreased 63% in PC3-shFRGR-4 tumors (p&lt;0.05) and the proportion of mice with lymph node metastasis was decreased from 93.3% to 50% (p&lt;0.01). The Ki67 immunohistochemistry showed a significant decrease of Ki67 positive cell percentage in PC3-sh-FGFR4 xenograft tumors (19.85%) compared to control group (35.93%, p&lt;0.001) indicating that the reduced primary tumor size was associated with the decreased cell proliferation. The orthotopic xenograft tumors produced with LNCaP-shFGFR4 showed 67.5% decreased weight compared to the tumors of control LNCaP-shV cells (p=0.0157). Our data indicated that FGFR4 can be a promising target in prostate cancer therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3271. doi:1538-7445.AM2012-3271
Abstract 328: The mitotic kinase Aurora-A promotes breast cancer metastases by inducing a CD24low/- basal-like stem cell phenotype
Authors: D'Assoro Antonino B. 
Published: August 2012

Abstract
In this study we demonstrate that gain of HER-2/Neu expression in human breast cancer xenografts harboring constitutive active Raf-1 induces Aurora-A over-expression that ultimately drives the transition from a luminal to a highly invasive basal-like phenotype. Over-expression of Aurora-A was induced by phosphorylation and protein stabilization by HER-2/Neu rather than increased Aurora-A gene expression. Moreover, Aurora-A stabilization induced an increase of HER-2/Neu expression in breast cancer cells defining a novel positive feed-back loop between Aurora-A and HER-2/Neu oncogenic signaling pathways. Basal-like tumor cells over-expressing Aurora-A and HER-2/Neu underwent to loss of CD24 surface receptor, expressed epithelial to mesenchymal transition (EMT) markers and acquired stem cell-like properties such as mammospheres formation and self-renewal. Pharmacologic inhibition of HER-2/Neu signaling pathway with lapatinib in cancer cells over-expressing Aurora-A demonstrated that Aurora-A was downstream to HER-2/Neu in the development of EMT and stemness. To determine whether Aurora-A induced a cancer stem cell-like phenotype through development of chromosomal instability and tumor cell phenotypic heterogeneity, we performed a SKY analysis on the breast cancer cell lines employed in this study. Our results demonstrate that invasive breast cancer cells harboring a basal-like phenotype did not develop higher chromosomal instability compared to luminal non-invasive cancer cells, signifying that Aurora-A has a direct effect on the development of basal-like breast cancer cells with stemness properties. Importantly, pharmacologic inhibition of Aurora-A kinase activity with the small molecule MLN8237 restored CD24 expression and reversed EMT leading to the re-establishment of a luminal phenotype and suppression of distant metastases. Our findings implicate for the first time the mitotic kinase Aurora-A in the development and maintenance of CD24 negative basal-like tumor stem cells responsible for breast cancer recurrence and progression. These observations have also important implications for the development of molecular targeted therapies to inhibit Aurora-A kinase activity and stemness improving the overall survival of breast cancer patients.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 328. doi:1538-7445.AM2012-328
Abstract 3283: Conditional Low-molecular-weight (LMW) cyclin E-induced mammary tumorigenesis
Authors: Akli Said , Van Pelt Carolyn S. , Wenbin Liu , Keyomarsi Khandan 
Published: August 2012

Abstract
Cyclin E overexpression and generation of hyperactive LMW cyclin E is oncogenic and has been implicated in a variety of human cancers. We have developed a doxycycline-inducible LMW cyclin E transgenic mouse model to determine the dependence of established mammary adenocarcinomas on continued expression of LMW cyclin E. Mice harboring this TetO-LMW-E transgene, referred to as TLMW, were mated to MMTV-rtTA transgenic mice (MTB) to yield bitransgenic MTB/TLMW mice. Morphological examination of hematoxylin and eosin-stained sections showed that the mammary glands of induced MTB/TLMW mice following four days of doxycycline induction were hyperplastic compared with uninduced MTB/ TLMW animals. Consistent with this, immunohistochemistry using antibodies specific for BrdU revealed a 2-fold increase in BrdU incorporation in the mammary epithelium of induced bitransgenic females compared with uninduced mice (17.1+/−5.3% versus 30.9+/−4.9%, P=0.03). This data indicate that LMW cyclin E overexpression in the mammary epithelium increased proliferation. To determine whether sustained induction of transgene expression would result in mammary tumorigenesis in MTB/TLMW mice, a cohort of 27 bitransgenic females was treated with doxycycline beginning at 6 weeks of age. 41% of these mice developed tumors within 2 years of starting doxycycline with kinetics (T1/2= 20 months) resembling that described for MMTV-LMW cyclin E mice. Similarly, the predominant type of tumor observed for the inducible LMW forms was glandular adenocarcinomas with adenosquamous differentiation. Lung metastases are known to arise in a fraction of MMTV-LMW cyclin E transgenic mice harboring primary mammary tumors. Accordingly, we sacrificed a subset of mammary tumor bearing MTB/TLMW mice and examined organs at necropsy for evidence of metastatic spread. 4 of 7 mice examined had lung metastasizes (57%). Control mice without dox never developed mammary tumors. To shed further light on the molecular mechanisms of mammary tumorigenesis caused by LMW-E expression, we conducted DNA microarray analyses. Using the R package limma, 641 probes out of 30,854 probes were identified as differentially regulated by at least 2-fold by LMW-E in the tumors when compared to the contralateral mammary glands. Gene Ontology analysis identified genes on this list that were involved in a variety of processes, including cell cycle control of chromosome replication, and DNA methylation and transcriptional repression signaling. Based on histology and gene expression profile of the resultant mammary tumors, we propose that the MTB/TLMW is a murine model of breast cancer resembling the human invasive basal/triple negative breast cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3283. doi:1538-7445.AM2012-3283