bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 700: Reduced CYLD expression is an independent prognostic factor in breast cancer: A possible link with RANKL-RANK signaling
Authors: Hayashi Mitsuhiro , Shinriki Satoru , Nakamura Takuya , Ibusuki Mutsuko , Sueyoshi Takanao , Ota Tomoko , Yamamoto Yutaka , Iwase Hirotaka , Ando Yukio 
Published: August 2012

Abstract
Background: Deubiquitinase CYLD plays an important role in tumorigenesis and immune response. RANKL-RANK system is required for osteoclast development and is also involved in bone metastasis in various malignancies while it has been also implicated in tumorigenesis, metastasis to distant organs other than bone in breast cancer through unknown mechanisms. Importantly, a recent study has reported that CYLD negatively regulated RANKL-RANK signaling in osteoclastogenesis. However, not only the role of CYLD but also its involvement in RANKL-RANK signaling in breast cancer is largely unknown. The purpose of this study was to elucidate clinical and biological significance of CYLD as well as its association with RANKL-RANK signaling in breast cancer. Materials and Methods: We assessed CYLD mRNA expression in 305 primary invasive breast cancer tissues including 26 matched-pair of tumoral and adjacent non-tumoral tissues, and 9 breast cancer cell lines as well as a nonmalignant breast epithelial cell line, HMEC by quantitative real-time RT-PCR. In addition, effect of CYLD knockdown by using siRNA on cell proliferation, migratory activity and the biological activity of RANKL in MDA-MB-231 cells. Results: Expression levels of CYLD mRNA were lower in tumoral lesion than the matched non-tumoral lesion in all of 26 cases. Our statistical analysis revealed that lower CYLD mRNA expression was significantly associated with reduced disease free survival rate (rog-rank, p = 0.025), irrespectively of breast cancer subtype. Multivariate analyses for disease free survival confirmed that low CYLD mRNA expression (Hazard ratio: HR 2.23, 95% CI 1.09 - 4.36, p = 0.029) in addition to estrogen receptor status (HR 0.15, 95% CI 0.05 - 0.44, p = 0.0003) and lymph node metastasis (HR 2.21, 95% CI 1.10 - 4.62, p = 0.026) were significant factors. Consistent with our clinical data, CYLD gene expression was reduced in all the breast cancer cell lines compared with HMEC cells. Repression of CYLD gene expression led to increased cell number in MDA-MB-231 cells. Interestingly, combination of CYLD knockdown and RANKL stimulation promoted cell migration while no change was observed in the setting of each alone. Conclusion: Downregulation of CYLD was common and an independent poor prognostic factor in invasive breast cancer. CYLD repression may promote breast cancer development and/ or progression in coordination with RANKL-RANK signaling.
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 700. doi:1538-7445.AM2012-700
Abstract 706: Ribonucleotide reductase small subunit M2 is a potential prognostic biomarker in human epidermal growth factor receptor 2- positive and triple negative breast cancers
Authors: Zhang Hang , Liu Xiyong , Deng Xutao , Liu Zheng , Warden Charles , Huang Yasheng , Bai Rui , Chen Huarong , Loera Sofia , Peng Jiaping , Zhou Lun , Ge Weiting , Hu Hanguang , Zhang Suzhan , Chu Peiguo , Zheng Shu , Yen Yun 
Published: August 2012

Abstract
Human epidermal growth factor receptor 2 (HER2)-positive breast cancer and triple negative breast cancer (TNBC) are two subtypes with poor outcome of all breast carcinomas. However, among all the patients in these two subtypes some still yielded relatively better prognosis. Ribonucleotide reductase (RR) small subunits M2 was associated with cancer cell invasiveness and metastasis, which prognoses poor survival in many cancers. Here we investigated whether RRM2 could serve as a prognostic biomarker for breast cancers, especially HER2- positive and TNBC subgroups. One hundred seventy five assessable breast cancers (40 cases were HER2-positive or TNBC) were collected from affiliated hospital of Zhejiang University (ZJU set); and six published microarray data sets (total 1154 cases, 63 cases were identified as HER2- positive or TNBC) with pathoclinical and follow- up information were employed for validation. In the ZJU set, the protein level of RRM2 was determined by IHC. It was indicated that RRM2 was positively correlated with cell proliferative marker Ki-67 (trends P=0.018) and stem/progenitor cell marker CD44+/CD24-/low (P=0.044). Multivariate COX model revealed that RRM2 was an independent prognostic biomarker for both OS (Hazard Ratio, HR=2.79, 95%Cl 1.21-6.75) and PFS (Hazard Ratio, HR=1.82, 95%Cl 1.02-3.26) for all the breast cancers. Moreover, in HER2-positive or TNBC breast cancers, RRM2-high was significantly associated with poor survival (P=0.025). Consequently, none of the patients with RRM2-low died of breast cancer in HER2-positive or TNBC subgroups. Furthermore, above findings were validated in the published data sets. The mRNA level of RRM2 evaluated significantly in highly-invasive subtypes including Luminal B, HER2-positive and TNBC in comparison with Luminal A and unclassified subgroups (P&lt;0.05). Overexpression of RRM2 was positively associated with poor differentiation (P&lt;0.05), advanced AJCC stage (P&lt;0.05) as well as poor survival (P&lt;0.05) in a dose-dependent manner. It was also revealed that high expression of RRM2 prognosed poor survival (HR=5.56, 95% CI 1.14-99.9) in HER2-positive breast cancers or TNBC. Moreover, the RRM2-high (protein or mRNA levels) was associated with better response to chemotherapy comparing to RRM2-low. Taken together, the above findings suggested that RRM2 was a potential biomarker for poor survival in breast cancer, especially in HER2- positive breast cancer and TNBC.
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 706. doi:1538-7445.AM2012-706
Abstract 708: Clinical significance of insulin-like growth factor 1 receptor (IGF1R) expression in pancreatic cancer
Authors: Hirakawa Toshiki , Yashiro Masakazu , Morisaki Tamami , Aomatsu Naoki , Fukuoka Tatsunari , Yoshii Mami , Hasegawa Tsuyoshi , Matsuoka Junko , Nagahara Hisashi , Kimura Kenjiro , Noda Eiji , Amano Ryosuke , Kubo Naoshi , Tanaka Hiroaki , Muguruma Kazuya , Yamada Nobuya , Maeda Kiyoshi , Nakata Bunzo , Ohira Masaichi , Hirakawa Kosei 
Published: August 2012

Abstract
Introduction: Insulin like growth factor 1 receptor (IGF1R) reported to be overexpressed in some types of human carcinomas, including lung cancer, and breast cancer. However no report of the significance of IGF1R expression was available in pancreatic cancer. Most patients with pancreatic cancer die within 2 years at diagnosis even after curative resection. The objective of this study was to evaluate the significance of IGF1R expression in human pancreatic cancer. Material and Methods: A total 122 patients with pancreatic cancer was studied. Paraffin embedded species of pancreatic cancers were stained with antibodies against IGF1R. IGF1R expression was evaluated by intensity of staining. Semi-quantitative scores of zero (no staining), 1+ (weak staining), 2+ (moderate staining), or 3+ (strong staining) were assigned to each sample based on the intensity of staining. Scores of 2+ or 3+ were defined as IGF1R-positive staining. The association between IGF1R expression and clinicopathologic variables was examined. Results: IGF1R expression was positive in 50 (40.1%) of 122 cases. Kaplan-Meier analysis demonstrated the prognosis of IGF1R-positive patients was significantly (p=0.0076) poorer than that of IGF1R-negative, and median survival time of patients with IGF1R-positive was 37.3 months, while that of patients with IGF1R-negative samples was 60.8 months. There were no associations between IGF1R overexpression and clinicopathological factors, including tumor location, tumor size, tumor differentiation, and T/N categories according to the International Union against Cancer. Multivariate analysis showed IGF1R expression and lymph node metastasis were independent predictors of worse prognosis in pancreatic cancer. Conclusion: The expression of IGF1R might be a novel predictive prognostic marker for patients with pancreatic cancers.
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 708. doi:1538-7445.AM2012-708
Abstract 710: Genetic identification of Annexin-A8 as a therapeutic target and prognostic marker for pancreatic cancer
Authors: Pimiento Jose M. , Chen Dung-Tsa , Centeno Barbara , Husein Kazim , Wang Chen , Zhang Anying , Malafa Mokenge P. 
Published: August 2012

Abstract
Introduction: We previously have identified histologic grade as an important prognostic factor for outcomes in early stage pancreatic cancer following surgical resection. In this study we investigated genes associated with histologic differentiation in early stage pancreatic cancer and their biologic relevance. Methods: We studied the gene expression profile of 61 resected primary pancreatic cancers through analysis of a custom Affymetrix microarray. Significance analysis of microarrays was used to select genes associated with histologic grade. Following identification of these genes we focused on Annexin A8 (A-A8) for further mechanistic studies. We analyzed the relationship of protein expression, tumor grade and outcomes using immunohistochemistry (IHC) in a tissue microarray (TMA) of 91 resected pancreatic cancer patients. Mechanistic studies of A-Aa8 included gene silencing using SiRNA and measurement of the cellular effects by migration, proliferation and colony formation assays. Statistical analysis was performed with Fisher's exact test, T-test and ANOVA as indicated and Cox modeling was used for survival analysis. Results: We identified 34 genes that were downregulated, and 15 genes that were upregulated as tumor grade progressed from well to moderate to poorly differentiated. Then we focused on A-A8 induction in this data set, with a 5.9 fold increased A-A8 expression from low to high grade tumors (r=0.31;p=0.019), the immunohistochemical expression of A-A8 was correlated with tumor grade in a human pancreatic tumor tissue microarray (r=0.26;p=0.014); finally high expression of A-A8 is associated with a significant decrease in 5 year survival following resection of stage I and II human pancreatic cancer by Cox model (p=0.049). Western blotting confirmed increased expression of A-A8 in human pancreatic cancer cell lines. Knockdown of A-A8 in human pancreatic cancer BXPC-3 cells with SiRNA resulted in significant decrease in cell viability (p&lt;0.001), cell migration (2.5 vs. 0.9mm at 96h;p=0.048), and colony formation (p=0.007). Conclusions: Studying the expression profiles of primary pancreatic cancer analyzed on Affymetrix microarrays, we identified A-A8 as a gene associated with pancreatic cancer differentiation. Downregulation of A-A8 by RNA interference in pancreatic cancer cells results in growth and migration inhibition. Finally we show that high A-A8 expression is associated with poor disease outcome in pancreatic cancer patients. These results support further investigation of Annexin A8 as a potential therapeutic target and prognostic marker for human pancreatic 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 710. doi:1538-7445.AM2012-710
Abstract 713: GLRX3, a secretory biomarker of pancreatic cancer based on pancreatic cancer stem cell characteristics
Authors: Kim Sun A , Park Soo Bin , Song Si Young 
Published: August 2012

Abstract
A. BACKGROUND/AIMS Pancreatic cancer is one of the most lethal diseases, which is difficult to diagnose and resistant to conventional treatment such as chemotherapy and radiotherapy. Cancer stem cells are involved in carcinogenesis, cancer progression and recurrence. We tried to find secretory biomarkers associated with pancreatic cancer stem cells using proteomic analysis. B. METHODS Pancreatic cancer stem like cells were enriched using sphere culture method. Proteomic analysis was done with secreted protein in culture medium of sphere and adherent cells. Identified proteins were confirmed with Western blot and immunohistochemical staining. ELISA was done to detect blood levels of validated proteins in pancreatic cancer patients. shRNA for GLRX3 was used to study the function of GLRX3 in pancreatic cancer. C. RESULTS Proteomic analysis showed that total of 55 protein spots from sphere cells were increased compared to adherent cells. Among them, 41 spots were identified using MALDI-TOF. Proteins known to be associated with cancer stem cells such as HSP90AB1, ALDH, and vimentin were expressed in sphere cells. The overexpression of 5 proteins including GLRX3 was confirmed by western blot in sphere cells and patient serum. Immunohistochemical staining in TMA of human pancreatic cancer tissue revealed that GLRX3 was detected in 15 (57.7%) out of 25 pancreatic cancer tissues regardless of cancer stages. To examine GLRX3 involvement in pancreatic carcinogenesis, we used shRNA to generate GLRX3-knockdown cells. Compared to the mock, knockdown of GLRX3 in human pancreatic cancer cell lines decreased in vitro proliferation, clonogenecity, and sphere formation. GLRX3 knockdown reverses epithelial-mesenchymal transition in pancreatic cancer cells. ELISA showed patients with unresectable pancreatic cancer and high blood GLRX3 levels had poor survival compared to those with low blood GLRX3 levels (155 days vs 463 days, p-value &lt;0.000). D. CONCLUSION This study shows that secretory protein GLRX3 may be a useful prognostic marker in pancreatic cancer. More researches are needed to find their roles in cancer stem cells and clinical implication.
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 713. doi:1538-7445.AM2012-713
Abstract 717: Aldo-keto reductase family 1B10 (AKR1B10): Novel biomarker and molecular target of hepatocellular carcinoma
Authors: Bai Han , Zhang Wanying , Li Haonan , Matkowskyj Kristina A. , Yang Allison L. , Liao Jie , Yang Guang-Yu 
Published: August 2012

Abstract
Aldo -keto reductase family 1B10 (AKR1B10) is a member of aldo-keto reducase (AKR) superfamily with biologic functions involved in osmolytic regulation, carbonyl detoxification, and carcinogenesis. Overexpression of AKR1B10 is identified in tumors of liver, lung, cervix and pancreas. However, the functional role of AKR1B10 in cancer remains unclear. In the present studies, we determined the expression and function of AKR1B10 in human hepatocellular carcinoma (HCC) using ex vivo HCC tissues and HCC cell lines. Thirty nine of HCC surgical resected specimens have been analyzed for the expression of AKR1B10 using immunohistochemistry method and revealed that 92.3% (36/39) of HCC expressed AKR1B10, including 100% (10/10) well differentiated, 95.2% (20/21) moderate differentiated, and 75% (6/8) poor differentiated HCC. No expression was observed in the adjacent morphologically normal liver. Expression of AKR1B10 in HCC confined to the cytoplasm. In order to investigate the oncogenic role of AKR1B10 in HCC, silence or inhibition of AKR1B10 in HCC cells (Huh 7) was approached using lentivirus mediated shRNA method or AKR1B10 inhibitor epalrestat. The results showed that the expression of AKR1B10 protein was significantly decreased in the AKR1B10 silenced cell lines with three different shRNA sequences using western blot approach. Knockdown AKR1B10 resulted in a significant inhibition of Huh 7 cell proliferation/tumorigenesis by colony formation assay and showed the average clone number was 174±25 of pGIPZ vector control Huh7 cell, and 86±32 of the AKR1B10 shRNA silencing cells (p&lt;0.05). The average clone size was also significantly decreased in AKR1B10 shRNA silencing cells. A slightly increased cell apoptosis was found in AKR1B10 silenced cells as detected with both caspase-3 immunohistochemistry (IHC) and Annexin V flow cytometry. Furthermore, AKR1B10 silenced cells or pGIPZ vector control Huh7 cells pretreated with AKR1B10 inhibitor epalrestat significantly increased sensitivity to doxorubicin and 4-hydroxy Hexenal-induced cell death as measured with Annexin V flow cytometry. Reduction of GTP-KRAS activity and its downstream MAPK signals were observed in AKR1B10 silencing cells as measured by western bolt approach. Our results indicated that AKR1B10 is involved in the process of cell proliferation, apoptosis, and Kras activation, and further imply that AKR1B10 is a novel biomarker and crucial molecular target of HCC.
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 717. doi:1538-7445.AM2012-717
Abstract 74: The role of epithelial splicing regulatory protein 1 in ovarian cancer
Authors: Yuan Chengfu , He Xiaolong 
Published: August 2012

Abstract
Epithelial splicing regulatory protein 1 (ESRP1) is a recently identified cell type-specific splicing factor which has been shown to control an alternative splicing network associated with the epithelial-mesenchymal transition (EMT). Whether ESRP1 play any roles in the development and progression of epithelial ovarian cancer is unknown. To answer this question, we first examined its expression by RT-PCR in a panel of cultured cells including normal ovarian surface epithelial cells (HOSE), immortalized OSE (IOSE398 and IOSE120T) as well as ovarian cancer cell lines A2780, OVCA3, OVCAR5, OVCAR8 and SKOV3. We found that ESRP1was expressed at low levels in HOSE, IOSEs and A2780 cells, at intermediate levels in OVCAR8 and SKOV3 cells and at high levels in OVCAR3 and OVCAR5 cells. Examination of ESRP1 target gene FGFR2 showed that cells with low to intermediate levels of ESRP1 exclusively expressed mesenchymal form of FGFR2 (FGFR2IIIc) while cells with high levels of ESRP1 exclusively expressed epithelial form of FGFR2 (FGFR2IIIb). These results suggest two notions: 1) the switch of FGFR2 splicing occurs in an all-or-none format dependent on the threshold of ESRP1 expression levels; 2) normal ovarian surface epithelial cells may have mesenchymal properties. We also examined the expression of ESRP1 in seven pairs of matched ovarian tumor and adjacent normal tissues and found that ovarian tumors expressed higher levels of ESRP1 than normal tissues in all pairs. In the attempt to clone the entire coding sequence of ESRP1 from SKOV3 cells, we have identified 11 ESRP1 splice variants (SVs). Ectopic expression of four ESRP1 SVs in 293T and A2780 cells, which both have very low levels of endogenous ESRP1, triggered profound apoptosis. The two SVs with exons 14 and 15 skipped appeared more cytotoxic than full-length ESRP1. We are currently investigating the effects of knockdown of ESRP1 on the transformation properties of ovarian cancer 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 74. doi:1538-7445.AM2012-74
Abstract 75: Identification and characterization of novel MELK (maternal embryonic leucine zipper kinase) substrates in breast cancer cells
Authors: Chung Suyoun , Nakamura Yusuke 
Published: August 2012

Abstract
Breast cancer is the most common malignancy among women worldwide. Treatment for breast cancer acting on molecular targets in critical signaling pathways in cancer cells have successfully reduced the motility rate in a subset of breast cancer patients, but a significant portion of patients cannot expect to have benefits from these treatments. Therefore, development of new therapeutics for those patients urgently required. Toward the goal of cancer-specific drug development, we previously identified MELK (maternal embryonic leucine zipper kinase) as a target for development of novel breast cancer therapy through genome-wide expression profile analysis. MELK was over-expressed in the great majority of breast cancer cells, particularly in TN breast cancer cells, and knockdown of MELK significantly suppressed the growth of breast cancer cells. Concordantly, the cells, which were transiently induced MELK, promoted its proliferation. Furthermore, cells in which over-expression of MELK were likely to increase the proportion of breast cancer-initiating stem cells more than those transfected with mock vector or kinase-dead MELK mutant plasmid. Although several substrates of MELK were reported, we are still not fully understood how MELK contributes to breast cancer development or progression. To elucidate the MELK signaling pathway in breast cancer cells, we examined 2D-PAGE and mass spectrometric analysis. Finally, we successfully identified two novel substrates of MELK. We confirmed the phosphorylation of these substrates by MELK via in vitro kinase assay and identified phosphorylation sites on each substrate by mass spectrometry. These results provide new information for better understanding of the biological role of MELK.
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 75. doi:1538-7445.AM2012-75