bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5295: Dysregulated miR-363 affects head and neck cancer invasion and metastasis by targeting PDPN gene.
Authors: Chen Wantao
Published: August 2014

Head and neck squamous cell carcinoma (HNSCC) is characterised by an elevated capacity for cell invasion and lymph node metastasis. The cause remains to be determined. Recent studies suggest that microRNAs can regulate the evolution of malignant behaviours by regulating multiple target genes. In this study, we first confirmed that miR-363 is down-regulated in HNSCC tissues with lymph node metastasis and cell lines with highly invasive capacity. We also found that podoplanin (PDPN) is up-regulated in the metastatic HNSCC tissues, and ectopic over-expression of miR-363 suppressed the metastatic behaviour of HNSCC cells by directly targeting PDPN. These data support a key role of miR-363-PDPN in head and neck cancer metastasis by regulating cellular invasion and migration.

Citation Format: Wantao Chen. Dysregulated miR-363 affects head and neck cancer invasion and metastasis by targeting PDPN gene. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5295. doi:10.1158/1538-7445.AM2013-5295
Abstract 5298: Two distinctive single nucleotide polymorphisms determine liver metastases responses to docetaxel plus thiotepa for metastatic breast cancer patients.
Authors: Yu Jing , Dong Ningning , Yan Ying , Shao Bin , Di Lijun , Song Guohong , Che Li , Jia Jun , Jiang Hanfang , Liang Xu , Zhu Yulin , Wang Chaoying , Zahng Jie , Zhu Budong , Zhou Xinna , Wang Xiaoli , Yang Huabing , Ren Jun , Lyerly Herbert Kim
Published: August 2014

The successful predictively treatment of metastatic breast cancer (MBC) remains a major challenge. Until now there is no definite marker to distinguish responder from non-responder and more and more important, metastasis occurred within different organs lead to serious variations of overall survival. In this study, paclitaxel and anthracyclines pretreated153 patients with MBC received 606 cycles of docetaxel plus thiotepa. 65 of 153 patients were concurrently diagnosed as MBC involved liver metastasis (LM). Among 153 patients, the response were 1 complete response (CR) (0.7%), 25 partial responses (PR) (16.3%), 73 stable diseases (SD) (47.7%), 46 progressive diseases (PD) (30.1%) and 8 unevaluable(5.2%). For LM patients, the localized liver response were 2 CR (3.1%), 20 PR (30.8%), 16 SD (24.6%), 22 PD (33.8%),5 unevaluable (7.7%). Median PFS and OS for both entire and LM subgroup were 6.5 (95% CI, 5.6 to 7.4) versus 4.2 (95%CI, 2.3-6.1), and 20.0 (95% CI, 15.6 to 24.4) versus 14.2 (95%CI, 9.5-18.9) months respectively. 79 SNPs in CYP450, whose minor allele frequency were ≥ 10% were genotyped. There was no significant difference of SNPs in therapeutic responses, PFS or OS. Of importance, there were two distinctive SNPs, rs2277119 and rs4646487 *2/*3 alleles, which tended to have the better liver metastases response than *1 when choosing a false discovery rate as 12%. It seemed that docetaxel plus thiotepa might be regarded as an active specific regimen for MBC with liver metastasis. The generation of conceptually specific chemotherapy targeting the organ with cancer metastasis should be considered in the future.

Citation Format: Jing Yu, Ningning Dong, Ying Yan, Bin Shao, Lijun Di, Guohong Song, Li Che, Jun Jia, Hanfang Jiang, Xu Liang, Yulin Zhu, Chaoying Wang, Jie Zahng, Budong Zhu, Xinna Zhou, Xiaoli Wang, Huabing Yang, Jun Ren, Herbert Kim Lyerly. Two distinctive single nucleotide polymorphisms determine liver metastases responses to docetaxel plus thiotepa for metastatic breast cancer patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5298. doi:10.1158/1538-7445.AM2013-5298

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5310: Activation of FGFR3 by genomic fusion in urothelial carcinoma of the bladder.
Authors: Williams Sarah V. , Hurst Carolyn D. , Knowles Margaret A.
Published: August 2014

FGFR3 is frequently activated by mutation in urothelial carcinoma of the bladder. We demonstrate here a further method of activation, by gene fusion following chromosomal re-arrangement. Four fusions with transforming acid coiled-coil 3 (TACC3) resulting from 4p16.3 re-arrangement were identified. A fifth fusion following t(4;7) was with BAI1-associated protein 2-like 1 (BAIAP2L1), also known as IRTKS (Insulin Receptor Kinase Substrate) on chromosome 7. The FGFR3-TACC3 fusions involve four different genomic re-arrangements resulting in three different transcripts. The fusion proteins show constitutive phosphorylation, activation of the MAPK pathway and transform NIH-3T3 cells. All have lost the final exon of FGFR3, which includes the PLCγ1 binding site. Thus PLCγ1 pathway activation and known PLCγ1-induced effects in normal urothelial cells were not induced. The FGFR3-BAIAP2L1 fusion retains most of BAIAP2L1 including the IRSp53/MIM domain (IMD) that mediates actin binding and Rac interaction, while the FGFR3-TACC3 fusions all retain the TACC domain that mediates microtubule binding. As cells containing these fusion genes are highly sensitive to FGFR-selective agents, detection of fusion genes may be useful in selecting patients for targeted therapy.

Citation Format: Sarah V. Williams, Carolyn D. Hurst, Margaret A. Knowles. Activation of FGFR3 by genomic fusion in urothelial carcinoma of the bladder. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5310. doi:10.1158/1538-7445.AM2013-5310
Abstract 5314: Calcium mediated growth control in normal and transformed colon cells: A possible role of the calcium sensing receptor.
Authors: Aggarwal Abhishek , Höbaus Julia , Fetahu Irfete Sh. , Mesteri Ildikó , Kállay Enikö
Published: August 2014

Introduction: Calcium (Ca2+) plays an important role in regulating proliferation of colonic epithelium. Epidemiological studies suggest an inverse correlation between dietary Ca2+ intake and colorectal cancer (CRC) risk. However in CRC, transformed cells lose their sensitivity to the tumour inhibiting effects of Ca2+. We have shown that the extracellular calcium-sensing receptor (CaSR) expression is lost in CRC and therefore hypothesized that the CaSR mediates the effect of Ca2+ in regulating proliferation.

Methods: We investigated the expression of the CaSR and of a cluster of genes responsible for ‘triggering off’ proliferation (CDT1, CDC6, CDC45, MCM2-7, Geminin, known as replication licensing genes) at mRNA level by qRT-PCR in CRC samples and in their respective ‘normal’ adjacent mucosa (n=57). To study the effect of Ca2+ in normal colon physiology, we fed male and female C57BL/6 mice with a diet containing high (0.9%) or low (0.1%) levels of Ca2+ and assessed the influence of dietary Ca2+ on proliferation (immunohistochemistry staining and mRNA expression) and CaSR mRNA expression.

Results and Discussion: In tumours we found the expression of CaSR significantly downregulated when compared with respective adjacent mucosa (p<0.001). All genes involved in replication licensing are significantly higher expressed (p<0.001) in these samples. Our data suggest that the loss of CaSR gives the tumour cells growth advantage. In the tumour samples we found negative correlation between CaSR and licensing factors (CDC45 (p<0.05, ρ=−0.265) and MCM6 (p<0.05, ρ=−0.262) reaching significance). Surprisingly, CaSR showed positive correlation with the licensing factors in the ‘normal’ mucosa (CDT1 (p<0.05, ρ=0.269), MCM2 (p<0.003, ρ=0.382) and MCM5 (p<0.001, ρ=0.490) reaching significance). These results are similar to our observation in the mouse study, where low dietary Ca2+ enlarged the proliferative zone of the crypts with no significant change in CaSR expression.

Conclusion: We conclude that the CaSR might regulate the calcium-dependent inhibition of proliferation in normal colonocytes. Upregulation of CaSR is probably a defence mechanism of the normal colon to control epithelial growth and counteract hyper-proliferative signals. This control mechanism is lost in cancer where CaSR expression is downregulated.

Citation Format: Abhishek Aggarwal, Julia Höbaus, Irfete Sh. Fetahu, Ildikó Mesteri, Enikö Kállay. Calcium mediated growth control in normal and transformed colon cells: A possible role of the calcium sensing receptor. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5314. doi:10.1158/1538-7445.AM2013-5314
Abstract 5317: Evidence for intracellular calcium-regulated secretion in gastrointestinal stromal tumor.
Authors: Berglund Erik , Berglund David , Akcakaya Pinar , Ghaderi Mehran , Dare Elisabetta , Berggren Per-Olof , Aspinwall Craig A. , Lui Weng-Onn , Zedenius Jan , Larsson Catharina , Bränström Robert
Published: August 2014

Gastrointestinal stromal tumors (GISTs) are considered to originate from the electrically active pacemaker cells of the gut. Despite the presence of synaptic like vesicles and proteins involved in exocytosis and secretory response in other cell types, it still remains unclear if GIST cells have a regulated secretion. A GIST tumor cell line (GIST882) was used as a model cell system, and stimulus-secretion coupling was investigated performing measurements of intracellular Ca2+, confocal microscopy, flow cytometry and an ATP-based assay to measure secretory response. We demonstrate that GIST cells have an intracellular signaling pathway downstream regulating cell secretion observed as an ATP release. Cell membrane integrity was preserved showing that ATP is released and not related to cell membrane rupture or cell death. The stimulus-secretion signal transduction is, at least partly, dependent on Ca2+ influx, since exclusion of extracellular Ca2+ diminishes the secretory response. We conclude that measurements of the secretory response in GIST by using ATP as a marker may be a useful tool in dissecting the signal transduction pathway, identification of secretory substances and in the development of drugs.

Citation Format: Erik Berglund, David Berglund, Pinar Akcakaya, Mehran Ghaderi, Elisabetta Dare, Per-Olof Berggren, Craig A. Aspinwall, Weng-Onn Lui, Jan Zedenius, Catharina Larsson, Robert Bränström. Evidence for intracellular calcium-regulated secretion in gastrointestinal stromal tumor. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5317. doi:10.1158/1538-7445.AM2013-5317

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5318: The breast epithelial stem cell line D492 overexpressing miR200c-141 is resistant to mesenchymal transition.
Authors: Hilmarsdottir Bylgja , Sigurdsson Valgardur , Bergthorsson Jon T. , Sigridur Franzdottir R. , Magnus Magnusson K. , Gudjonsson T
Published: August 2014

MicroRNAs (miRs) are highly conserved, small RNA molecules that regulate key biological processes. miRs can act as tumor suppressors or oncogenes and their expression is often deregulated in cancers. The miR200 family is found in two clusters at chromosome 1 (miR200ba-429) and 12 (miR200c-141). Downregulation of the miR200 family has been linked to epithelial to mesenchymal transition (EMT) and recent studies show that members of this family are regulators of epithelial integrity in many tissues. We have recently shown that D492, a breast epithelial cell line with stem cell properties which generates branching structures when cultured in reconstituted basement membrane (rBM), undergoes EMT in coculture with endothelial cells. This is evidenced by formation of a stable spindle-like phenotype, suppression of keratin expression and cadherin switch from E- to N-cadherin (Sigurdsson et al. PLoS ONE 2011). In this study we have mapped the miRNA expressional changes in D492 compared to its mesenchymal-derivative D492M. Among the most profound changes were suppression of the miR200 family. Treatment of D492M with the demethylating agent 5-aza-2′-deoxycytidine reversed the EMT phenotype and reestablished E-cadherin expression. Bisulfite sequencing showed that the promoter region of miR200c-141 was methylated in D492M but not in D492. No methylation was seen in the promoter regions of E-cadherin or miR200ba-429 in either cell line. Overexpression of miR200c-141 in D492M reversed the EMT phenotype resulting in loss of N-cadherin and Snail expression and gain of E-cadherin and EpCAM expression. Interestingly, overexpression of miR200c-141 in D492M failed to reverse keratin expression back to an epithelial phenotype. When cultured in rBM D492M-200c-141 cells form irregular branching and showed no EMT phenotype. Interestingly, overexpression of miR200c-141 in D492M and D492 inhibits endothelial induced mesenchymal transition in rBM. This indicates that silencing of the miR-200c-141 with promoter methylation is associated with endothelial induced-EMT. In conclusion, our data suggests that overexpression of miR-200c-141 in D492 and D492M inhibits and reverses EMT, respectively, and this support the view that miR200c-141 may be a potent tumor suppressor and important regulator of epithelial integrity in the human breast.

Citation Format: Bylgja Hilmarsdottir, Valgardur Sigurdsson, Jon T. Bergthorsson, Franzdottir R. Sigridur, Magnusson K. Magnus, T Gudjonsson. The breast epithelial stem cell line D492 overexpressing miR200c-141 is resistant to mesenchymal transition. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5318. doi:10.1158/1538-7445.AM2013-5318
Abstract 532: Differential expression of FGFR-1 and -2 in hormone responsive and resistant murine mammary carcinomas.
Authors: Sahores Ana , Guillardoy Tomás , Wargon Victoria , Lanari Claudia , Lamb Caroline
Published: August 2014

Fibroblast growth factor (FGF) receptors (FGFRs) are dysregulated in a number of developmental and neoplastic conditions. In human breast cancer samples we have previously demonstrated a significant association between FGFR2 expression and estrogen receptors as well as a positive correlation of FGFR1 and high histological grade. Most breast carcinomas that express hormone receptors respond initially to an endocrine therapy, but over time, they develop resistance (acquired hormone resistance). Others fail to respond from the beginning (constitutive resistance). Using a breast cancer mouse model, we have previously demonstrated that antiprogestin-responsive tumors (C4-HI) show a higher expression level of progesterone receptor isoform A than PR isoform B (PRB), while tumors with constitutive (C4-2-HI) or acquired resistance (C4-HIR) to antiprogestins, display a higher expression level of PRB. Moreover, we have demonstrated, in an antiprogestin-responsive tumor that FGFR2 activated by FGF2 released by the stromal compartment participate in tumor growth activating PR. The aim of this study is to investigate the expression of FGF2, FGFR1, FGFR2 in C4-HI, C4-HIR and C4-2-HI tumor variants. By immunohistochemistry and western blot we observed a decrease in FGFR2 expression for C4-2-HI (p<0.001) and C4-HIR (p<0.01) as compared to C4-HI. Moreover, we found an increased expression of FGF2 and FGFR1 by western blot (p<0.05) and immunohistochemistry in the epithelium of the resistant tumor variants. Similar results were obtained with another family of antiprogestin responsive and unresponsive tumors (59-HI and 59-2-HI). These data suggest that in this mammary tumor model hormone resistance may be characterized by a switch from paracrine to autocrine FGF2 signaling together with a decrease in FGFR2 and an increase in FGFR1. These results contribute to understand the role of the FGFR pathway in hormone resistance and support the use of FGFR inhibitors combined with hormonal therapy to delay the onset of hormone resistance.

Citation Format: Ana Sahores, Tomás Guillardoy, Victoria Wargon, Claudia Lanari, Caroline Lamb. Differential expression of FGFR-1 and -2 in hormone responsive and resistant murine mammary carcinomas. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 532. doi:10.1158/1538-7445.AM2013-532
Abstract 5324: MicroRNA-23b contributes to stem cell overpopulation in colon carcinomas.
Authors: Viswanathan Vignesh , Opdenaker Lynn , Boman Bruce
Published: August 2014

How stem cell (SC) overpopulation drives colorectal cancer (CRC) development and growth is unclear. Because microRNAs (miRNAs) normally regulate SC populations, we reasoned that aberrant expression of these miRNAs are involved in CRC. Our goal was to identify such miRNAs. We hypothesized that there is a miRNA expression signature for SCs in the normal colonic crypt and that these miRNAs are aberrantly expressed in colon cancer SCs (CSCs). Accordingly, we used a 368-gene microarray chip to identify miRNAs that show differential expression in the crypt bottom (the SC niche) vs. the crypt top. A subset of 16 miRNAs were found to be significantly (p < 0.05) over- or under-represented in the crypt bottom. Four of the 16 - hsa-mir-206, hsa-mir-7, hsa-mir-25, hsa-mir-23b -were differentially expressed in CRC vs. normal colonic epithelium. Computational target prediction analysis indicated that miR-23b targets mRNAs encoding colon SC-specific markers (ALDH1, LGR5 and LRIG1). MicroRNA-23b was down-regulated in the crypt bottom and upregulated in ALDH+ (vs. ALDH-) cells isolated from HT29 and SW480 CRC lines. Manipulation of miR-23b levels (using a precursor construct) in these cell lines decreased proliferation by causing G0/G1 cell-cycle growth arrest (vice versa with the anti-miR-23b construct). Decreasing miR-23b levels in HT29 cells inhibited their colony and colonosphere-formation ability, indicating that miR-23b has a role in CSC self-renewal. Thus, miR-23b appears to be expressed at low levels in normal colonic SCs; when upregulated in CSC, it appears to contribute to CSC overpopulation and CRC development. Targeting miR-23b may represent an innovative approach to target CSC overpopulation in CRC treatment.

Citation Format: Vignesh Viswanathan, Lynn Opdenaker, Bruce Boman. MicroRNA-23b contributes to stem cell overpopulation in colon carcinomas. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5324. doi:10.1158/1538-7445.AM2013-5324
Abstract 5328: MicroRNA-181a suppresses salivary adenoid cystic carcinoma metastasis by targeting the ERK/Slug pathway.
Authors: He Qianting , Zhou Xiaofeng , Li Su , Chen Zhujian , Chen Dan , Cai Yuchen , Liu Zhonghua , Zhao Tingting , Wang Anxun
Published: August 2014

MicroRNA deregulation plays a major role in the initiation and progression of many cancers. The objective of this study is to evaluate the role of miR-181a in the metastasis of salivary adenoid cystic carcinoma (SACC). We identified a panel of differentially expressed microRNAs in two pairs SACC cell lines with different metastatic potential using microarrays. Among these differentially expressed microRNAs, the down-regulation of miR-181a was further validated by quantitative RT-PCR. Functional analysis indicated that miR-181a inhibits SACC cell migration, invasion and proliferation in vitro and in vivo. Ectopic transfection of miR-181a inhibited the expression of NGF, VEGF, TGFβ2, ERK1/2, pERK1/2 and Slug in SACC-LM cells which had higher expression of these genes compared to SACC-83 cells; while inverse results were found in miR-181a LNA transfected SACC-83 cells. The direct targeting of miR-181a to the Slug and ERK2 mRNAs were confirmed by luciferase reporter gene assays. SiRNA-mediated knockdown of Slug, ERK2 led to reduced cell migration and invasion of SACC, and knockdown of ERK2 suppresses the expression of Slug. Taken together, these results suggest that microRNA deregulation (i.e., down-regulation of miR-181a) contributes to the metastasis of SACC. MiR-181a-mediated suppression of SACC metastasis may be achieved in part by directly targeting Slug and ERK2 and regulating ERK/Slug pathway.

Citation Format: Qianting He, Xiaofeng Zhou, Su Li, Zhujian Chen, Dan Chen, Yuchen Cai, Zhonghua Liu, Tingting Zhao, Anxun Wang. MicroRNA-181a suppresses salivary adenoid cystic carcinoma metastasis by targeting the ERK/Slug pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5328. doi:10.1158/1538-7445.AM2013-5328
Abstract 533: Differential expression of IGF-II in Vietnamese women in triple negative breast cancer is dependent on biallelic or monoallelic IGF-II with ApaI SNP.
Authors: Radhakrishnan Vinodh Kumar , Tan Qianwei , De León Marino A. , De León Daisy D.
Published: August 2014

Triple negative breast cancer (TNBC) is more likely to be advanced, poorly differentiated, larger, and present in women of younger age and of lower socioeconomic status. Moreover, TNBC patients have worse outcomes when treated with currently available adjuvant chemotherapy. We have demonstrated that Breast cancer (BC) tumors from AA patients express significantly higher levels of IGF-II compared to Caucasian women (CA), and show a higher activation of the IGF signaling pathways in tumors progression (Kalla et.al., 2010). Thus we designed a study in younger Vietnamese women where TNBC incidence is increasing. A total of 24 paired BC samples were analyzed by gDNA PCR for IGF-II Apa I polymorphism to correlate with IGF-II mRNA and protein levels. The levels of IGF-II correlated with the imprinting status, i.e., Biallelic (BA; n=14)>>, Monoallelic with SNP (MAS; n=5)>, and Monoallelic non-ApaI SNP (MA; n=5)>. Higher IGF-II levels were expressed in tumor as compared to normal BC tissue samples. The free proIGF-II protein and mRNA levels were higher in biallelic tumor tissues as compared to the proIGF-II levels detected in monoallelic tumor samples. The growth factor receptors; Insulin receptor A (IRA), Insulin receptor B (IRB), Vascular endothelial growth factor receptor (VEGFR), Estrogen receptor-1 (ESR1), Progesterone receptor (PGR) and Human epidermal growth factor receptor (ERBB2) levels were determined to show if IGF-II monoallelic or biallelic dosage regulated the receptor levels. We conclude that biallelic IGF-II expression in TNBC tumors results in higher IGF-II mRNA, protein and related receptors levels than in monoallelic samples.

Citation Format: Vinodh Kumar Radhakrishnan, Qianwei Tan, Marino A. De León, Daisy D. De León. Differential expression of IGF-II in Vietnamese women in triple negative breast cancer is dependent on biallelic or monoallelic IGF-II with ApaI SNP. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 533. doi:10.1158/1538-7445.AM2013-533
Abstract 5331: A novel function of p21 in inhibition of epithelial-mesenchymal transition through microRNAs.
Authors: Li Xiao L. , Hara Toshifumi , Choi Youngeun , Subramanian Murugan , Francis Princy , Bilke Sven , Walker Robert L. , Pineda Marbin , Yang Yu-an , Luo Ji , Wakefield Lalage M. , Park Ben H. , Brabletz Thomas , Chowdhury Dipanjan , Meltzer Paul S. , Lal Ashish
Published: August 2014

The tumor suppressor p21 inhibits cell proliferation during the stress response. However, p21 can also directly regulate gene expression by repressing specific transcription factors. Here, we identified p21-regulated miRNAs by sequencing small RNAs from HCT116-p21+/+ and HCT116-p21-/- cells. Three abundant clusters, miR-200b-200a-429, miR-200c-141 and miR-183-96-182 were down-regulated in p21-depleted HCT116 and MCF10A cells. Loss of p21 induced epithelial-mesenchymal transition (EMT) and enhanced migration and invasion in multiple model systems. Identification of genome-wide targets of the miR-183-96-182 cluster indicated that miR-183 and miR-96 repressed common targets, including SLUG, ZEB1, ITGB1 and KLF4, to inhibit EMT, migration and invasion. In turn, elevated ZEB1 levels in HCT116-p21-/- cells directly repressed miR-183-96-182 cluster transcription, revealing a feedback loop. Re-introduction of miR-200, miR-183 or miR-96 in HCT116-p21-/- cells inhibited migration and invasion. These novel findings suggest that coordinated down-regulation of three miRNA clusters upon loss of p21 in unstressed cells promotes EMT, migration and invasion.

Citation Format: Xiao L. Li, Toshifumi Hara, Youngeun Choi, Murugan Subramanian, Princy Francis, Sven Bilke, Robert L. Walker, Marbin Pineda, Yu-an Yang, Ji Luo, Lalage M. Wakefield, Ben H. Park, Thomas Brabletz, Dipanjan Chowdhury, Paul S. Meltzer, Ashish Lal. A novel function of p21 in inhibition of epithelial-mesenchymal transition through microRNAs. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5331. doi:10.1158/1538-7445.AM2013-5331
Abstract 5337: MicroRNA-30c targets cytoskeleton genes involved in breast cancer cell invasion.
Authors: Bockhorn Jessica , Yee Kathy , Chang Ya-Fang , Prat Aleix , Huo Dezheng , Nwachukwu Chika , Dalton Rachel , Huang Simo , Swanson Kaitlin E. , Perou Charles M. , Olufunmilayo Olufunmilayo I. , Clarke Michael F. , Liu Huiping , Greene Geoffrey
Published: August 2014

Metastasis remains a significant challenge in treating cancer. MicroRNAs have emerged as important epigenetic regulators of various cellular processes during cancer development and progression. The goal of this study was to characterize signaling pathways for miRNA biomarkers that regulate breast cancer metastasis. Here we show that human breast tumor biomarker miR-30c regulates invasion by targeting the cytoskeleton network genes encoding Twinfilin 1 (TWF1) and Vimentin (VIM). Both VIM and TWF1 have been shown to regulate epithelial-to-mesenchymal transition (EMT). Similar to TWF1, VIM also regulates F-actin formation, a key component of cellular transition to a more invasive mesenchymal phenotype. To further characterize the role of the TWF1 pathway in breast cancer, we found that IL-11 is an important target of TWF1 that regulates breast cancer cell invasion and STAT3 phosphorylation. This miR-30c VIM/TWF1-IL11-pSTAT3 pathway will expedite the development of targeting strategies to prevent and treat breast tumor progression.

Citation Format: Jessica Bockhorn, Kathy Yee, Ya-Fang Chang, Aleix Prat, Dezheng Huo, Chika Nwachukwu, Rachel Dalton, Simo Huang, Kaitlin E. Swanson, Charles M. Perou, Olufunmilayo I. Olufunmilayo, Michael F. Clarke, Huiping Liu, Geoffrey Greene. MicroRNA-30c targets cytoskeleton genes involved in breast cancer cell invasion. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5337. doi:10.1158/1538-7445.AM2013-5337
Abstract 5342: Role of microRNAs in the development of prostate cancer.
Authors: Kumar Parameet , Dobi Albert , Petrovics George , Mohamed Ahmed , Srivastava Shiv , Biswas Roopa
Published: August 2014

Prostate Cancer (CaP) is the most common non-cutaneous form of cancer in men. It is also the second leading cause of cancer mortality in men in the USA. Oncogenic activation of the ETS Related Gene (ERG) as a result of gene fusions is the most common genomic alteration in prostate cancer reported to date. ERG has a milieu of transcriptional targets that regulate genes involved in various cellular processes including oncogenesis, inflammation, cell invasion, and DNA damage. Moreover, knockdown (KD) of ERG in TMPRSS2-ERG expressing CaP cells (VCaP) suppresses the growth and invasiveness of VCaP cells and in xenograft models. ERG protein has been implicated as a key player in the progression from pre-invasive to invasive disease status of CaP. Therefore, understanding mechanisms by which ERG promotes CaP progression is essential for developing novel prognostic/therapeutic targets for CaP. Our analyses of miRNA expressions in the human TMPRSS2-ERG fusion harboring VCaP cells compared to ERG-suppressed (with siRNA against ERG) VCaP controls indicate altered expression of miRNAs. Interestingly, we find that ERG suppresses the expression of miR-449a, which has been shown to regulate proliferation of CaP cells. In silico analyses (IPA and Genomatix) of cellular pathways targeted by these miRs focus on regulatory networks that are indicative of EMT and include molecules like DES (desmin), PDGFRA (platelet-derived growth factor receptor, alpha polypeptide) and TGFBR1 (transforming growth factor beta receptor 1). These pathways are potential candidate therapeutic targets. These data form the basis of new insights into the miRNA alterations in the context of major oncogenic activation in prostate cancer. Moreover, these miRNAs will provide novel prognostic marker and therapeutic targets for prostate cancer and will ultimately define CaP associated miRNAs in the context of ERG positive and ERG negative prostate cancers.

Supported by: United States Military Cancer Institute [RB] and [SS].

Citation Format: Parameet Kumar, Albert Dobi, George Petrovics, Ahmed Mohamed, Shiv Srivastava, Roopa Biswas. Role of microRNAs in the development of prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5342. doi:10.1158/1538-7445.AM2013-5342
Abstract 5345: MicroRNA mediated regulation of a novel gene, MIEN1, in cancer metastasis.
Authors: Rajendiran Smrithi , Vishwanatha Jamboor K.
Published: August 2014

Migration and invasion enhancer gene 1(MIEN1), previously known as C17orf37, C35, ORB3, XTP4, RDX12, and MGC14832, is located in the 17q12 region of the human chromosome. MIEN1 expression is minimal to negligible in normal cells and tissues compared to its abundant expression in various cancers, including breast, prostate, oral and gastrointestinal carcinomas. MIEN1 works as a signaling molecule through the AKT/NF-κB pathway by up-regulating key proteases, matrix metalloproteinase - 9, urokinase - type plasminogen activator and vascular endothelial growth factor. We have shown that MIEN1 plays an important role in the migration and invasion of cancer through increased filopodia formation, hence promoting metastasis. The overall objective of this study is to identify the mechanisms that lead to the differential regulation of MIEN1 between normal and cancer cells. Our hypothesis is that MIEN1 is post-transcriptionally regulated by specific microRNA(s) that is (are) down-regulated in cancer, thus explaining its aberrant increased expression in cancer. MicroRNA expression profiling pattern predict several microRNAs that can directly bind to 3′UTR of MIEN1 mRNA. In silico analysis followed by northern blotting and real time PCR indicate a novel microRNA that regulates MIEN1 expression. This was further confirmed by decrease or increase in RNA and protein levels of MIEN1 as well as its downstream signaling effectors, upon microRNA mimic or inhibitor addition, respectively. Direct binding of the microRNA with MIEN1 mRNA was further confirmed by luciferase reporter assays, while the scratch wound healing and tumor invasion assays showed the functional significance of this microRNA on migration and invasion. Since the importance of MIEN1 as a key signaling molecule in cancer is well established, understanding the mechanism(s) involved in the regulation of MIEN1 will aid in designing novel and effective therapeutic strategies to treat cancer patients.

Citation Format: Smrithi Rajendiran, Jamboor K. Vishwanatha. MicroRNA mediated regulation of a novel gene, MIEN1, in cancer metastasis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5345. doi:10.1158/1538-7445.AM2013-5345
Abstract 5346: MiR-331-5p targets receptor EphA2 and attenuates tumor growth and migration in NSCLC.
Authors: Sukka-Ganesh Bhagyalaxmi , Mohammed Kamal A. , Kaye Frederic , Nasreen Najmunnisa
Published: August 2014

Introduction: Receptor EphA2 is over expressed in NSCLC and is associated with tumor progression and poor survival. MicroRNAs (miRs) are short, non-coding RNAs that post-transcriptionally regulate gene expression. Dysregulated expression of miRs has been observed in variety of malignancies. However, the role of miR-331-5p in NSCLC growth and migration is not known. We hypothesized that EphrinA1 treatment promotes miR-331-5p expression in NSCLC and miR-331 may inhibit tumor growth and invasion by targeting receptor EphA2.

Methods: The A-549 and H-23 NSCLC cells were treated with Ephrin-A1 for 24 hr and the miR-331-5p expression was measured by real-time PCR analysis. EphA2 protein expression was studied through Western blotting analysis. The cell proliferation was studied by WST-1 assay. The tumor formation and invasion was determined by 3-D matrigel and Wound healing assays respectively. Luciferase assay was performed to confirm miR-331-5p as one of the direct and specific target for EphA2.

Results: The miR-331-5p expression was inversely correlated with EphA2 expression. Transfection of NSCLC with precursor miR-331-5p inhibited proliferation, and migration of NSCLC cells. In addition the ability of tumor colony formation was significantly inhibited when the cells were transfected with miR-331-5p precursors. We have identified predicted target binding site of EphA2 in its 3’ UTR region by miR-331-5p. This was confirmed by Luciferase assay. Over-expression of miR-331-5p in NSCLC cells repressed the endogenous levels of EphA2 protein.

Conclusion: This study for the first time reports, that miR-331-5p directly regulates EphA2 mRNA and protein expression in NSCLC. MiR-331 directly binds to 3’ -UTR region of EphA2 and inhibits tumor growth and migration. Our findings also provide an insight into the regulation of EphA2 expression and suggest that miR-331-5p has the capacity to regulate signalling pathways critical for the development and progression of NSCLC.

Funding: RC1 from Florida Department Health, and VA Merit

Citation Format: Bhagyalaxmi Sukka-Ganesh, Kamal A. Mohammed, Frederic Kaye, Najmunnisa Nasreen. MiR-331-5p targets receptor EphA2 and attenuates tumor growth and migration in NSCLC. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5346. doi:10.1158/1538-7445.AM2013-5346
Abstract 5348: Characterization of the EZH2-MMSET histone methyltransferase regulatory axis in cancer.
Authors: Asangani Irfan A. , Ateeq Bushra , Cao Qi , Dodson Lois , Pandhi Mithil , Kunju Lakshmi P. , Mehra Rohit , Lonigro Robert J. , Siddiqui Javed , Palanisamy Nallasivam , Wu Yi-Mi , Cao Xuhong , Kim Jung H. , Iyer Mathew K. , Maher Christopher A. , Kumar-Sinha Chandan , Varambally Sooryanarayana , Chinnaiyan Arul M. , Zhao Meng , Qin Zhaohui S.
Published: August 2014

Histone methyltransferases (HMTases), as chromatin modifiers, regulate the transcriptomic landscape in normal development as well in diseases such as cancer. Here, we molecularly order two HMTases, EZH2 and MMSET that have established genetic links to oncogenesis. EZH2, which mediates histone H3K27 trimethylation and is associated with gene silencing, was shown to be coordinately expressed and function upstream of MMSET, which mediates H3K36 dimethylation and is associated with active transcription. EZH2 transcript expression levels were compared to all HMTases (n=51) in our compendium of RNA-seq data comprising 474 malignant and benign samples, representing 14 different tissue types. MMSET displayed the strongest correlation with EZH2 expression. Similarly, a meta-analysis of 1755 samples from 22 published microarray gene expression studies in Oncomine revealed co-expression of MMSET and EZH2 in 15 different cancers. We found that the EZH2-MMSET HMTase axis is coordinated by a microRNA network and that the oncogenic functions of EZH2 require MMSET activity. Together, these results suggest that the EZH2-MMSET HMTase axis coordinately functions as a master regulator of transcriptional repression, activation, and oncogenesis and may represent an attractive therapeutic target in cancer.

Citation Format: Irfan A. Asangani, Bushra Ateeq, Qi Cao, Lois Dodson, Mithil Pandhi, Lakshmi P. Kunju, Rohit Mehra, Robert J. Lonigro, Javed Siddiqui, Nallasivam Palanisamy, Yi-Mi Wu, Xuhong Cao, Jung H. Kim, Mathew K. Iyer, Christopher A. Maher, Chandan Kumar-Sinha, Sooryanarayana Varambally, Arul M. Chinnaiyan, Meng Zhao, Zhaohui S. Qin. Characterization of the EZH2-MMSET histone methyltransferase regulatory axis in cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5348. doi:10.1158/1538-7445.AM2013-5348
Abstract 535: IGF-I stimulates amino acid transporter xC- function to reduce intracellular ROS level and promote proliferation in human breast cancer cells.
Authors: Yang Yuzhe , Becker Marc A. , Yee Douglas
Published: August 2014

Our laboratory identified SLC7A11/xCT as an IGF-I-induced gene specifically regulated through the adaptor protein IRS-1 in breast cancer cells. xCT encodes the functional subunit of the heterodimeric plasma membrane transport system xC-, which is critical for the cellular uptake of cystine in exchange for intracellular glutamate. xC- transporter is involved in the regulation of proliferation, metastasis, and treatment resistance in cancer. To determine the role for xC- expression in IGF-mediated breast cancer cell biology, two classes of breast cancer cell lines were studied: estrogen receptor (ER) positive cell lines (IRS-1 activated) MCF-7, ZR-75-1, and T47D; basal-like cell lines (IRS-2 activated) MDA-MB-231, BT549, and HS578T. Significantly increased xCT mRNA expression was observed only in ER positive cell lines, which was eliminated by IRS-1 specific siRNA (25 nM). IGF-I increased xCT protein expression measured by immunblots and flow cytometry only in ER positive cells in an IRS-1 dependent manner. To measure the function of upregulated xC-, extracellular levels of glutamic acid and intracellular level of total glutathione were measured in MCF-7 and MDA-MB-231. IGF-I regulated xC- transporter function only in IRS-1 activated MCF-7 cells. In MCF-7 cells, IGF-I-stimulated monolayer and anchorage-independent growth were suppressed by treating cells with xCT shRNA or the xCT chemical inhibitor sulfasalazine (SASP, 0.1 mM). Direct measurement of reactive oxygen species (ROS) and levels of phospho-p38MAPkinase showed that IGF-I reduced cellular ROS level (induced by 10 Gy of irradiation or 1 ug/ml of mitomycin C), SASP (0.1 mM) reversed this IGF-I effect. Anchorage-independent growth assays suggested that inhibiting xC- function by 0.1 mM SASP sensitized the response of MCF-7 cells to anti-IGF-IR monoclonal antibody humanized EM164 (ImmunoGen, Inc.) and tyrosine kinase inhibitor NVP-AEW-541. These data suggest that IGF-I promotes the proliferation of ER positive breast cancer cells by regulating xC- transporter function in an IRS-1 dependent manner. Furthermore, IGF-I protects cells from ROS via upregulation of xC-. Our findings also imply that combination of xC- transporter inhibitor with anti-IGF-IR agents may have synergic therapeutic effects.

Citation Format: Yuzhe Yang, Marc A. Becker, Douglas Yee. IGF-I stimulates amino acid transporter xC- function to reduce intracellular ROS level and promote proliferation in human breast cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 535. doi:10.1158/1538-7445.AM2013-535
Abstract 5350: Enhancer of zeste homolog 2 (EZH2) promotes progression of cholangiocarcinoma by regulating cell cycle and apoptosis.
Authors: Nakagawa Shigeki , Okabe Hirohisa , Sakamoto Yasuo , Hayashi Hiromitsu , Hashimoto Daisuke , Kuroki Hideyuki , Imai Katsunori , Nitta Hidetoshi , Chikamoto Akira , Watanabe Masayuki , Beppu Toru , Baba Hideo
Published: August 2014

Background: Enhancer of zeste homolog 2 (EZH2) is a catalytic subunit of the polycomb repressive complex 2 (PRC2). When present in PRC2, EZH2 catalyzes trimethylation on lysine 27 residue of histone H3(H3K27Me3), resulting in epigenetic silencing of gene expression and cancer progression. Here, we investigated the expression and the function of EZH2 in intrahepatic and extrahepatic cholangiocarcinoma (ICC and ECC).

Methods: The influence of EZH2 on cell growth and apoptosis was assessed by knockdown experiments using siRNA in cholangiocarcinoma cell lines, RBE and TFK-1. Target genes of EZH2 in cholangiocarcinoma cell lines were searched by real time PCR. The clinical significance of EZH2 in 84 cholangiocarcinoma patients (ICC and ECC in 45 and 41, respectively) who underwent a curative surgery was examined by immunohistochemistry.

Results: In vitro analysis, a knockdown of EZH2 reduced cell growth and induced G1 arrest, and induced apoptosis confirmed by Annexin V staining and increasing sub-G1 population of two cholangiocarcinoma cell lines. Moreover, a knockdown of EZH2 increased the expression of p16INK4A and p27KIP1, in real time PCR. In immunohistochemical study, The upregulation of EZH2 was correlated with tumor diameter (P = 0.0103) in ICC, lymph node metastasis (P=0.0292) in ECC, and Ki67 index in both ICC (P=0.0364) and ECC (P=0.0017). In addition, EZH2 expression was correlated with the poor prognosis in both ICC (P=0.0447) and ECC (P=0.0227).

Conclusion: The current study demonstrates that the high expression of EZH2 results in acceleration of cell cycle and anti-apoptosis in vitro analysis, and is related to poor prognosis in patients with ICC or ECC by IHC study. These results suggest that EZH2 is a potential target for cholangiocarcinoma therapeutics.

Citation Format: Shigeki Nakagawa, Hirohisa Okabe, Yasuo Sakamoto, Hiromitsu Hayashi, Daisuke Hashimoto, Hideyuki Kuroki, Katsunori Imai, Hidetoshi Nitta, Akira Chikamoto, Masayuki Watanabe, Toru Beppu, Hideo Baba. Enhancer of zeste homolog 2 (EZH2) promotes progression of cholangiocarcinoma by regulating cell cycle and apoptosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5350. doi:10.1158/1538-7445.AM2013-5350
Abstract 5351: Phosphorylation of EZH2 at Thr487 is a marker for good prognosis in pancreatic cancer patients.
Authors: Wei Yongkun , Chang Shih-Shin , Wang Huamin , Xia Weiya , Hung Mien-Chie
Published: August 2014

EZH2 is the catalytic subunit of the polycomb repressive complex 2 (PRC2) and is involved in repressing gene expression through methylation of histone H3 on lysine 27 (H3K27). EZH2 is overexpressed in multiple tumor types and correlates with poor prognosis. We have previous shown that phosphorylation of EZH2 at Thr 487 by CDK1 inhibits its methyltransferase activity. However, the status of phosphorylation of EZH2 at Thr 487 (P-EZH2-T487) and its clinical implication in cancer patients have not been reported. We thus examined P-EZH2-T487 by immunohistochemistry and its association with prognosis in pancreatic cancer patients. We found that P-EZH2-T487 can be seen in 61% (47/77) of pancreatic cancer. In univariate survival analysis, patients with high P-EZH2-T487 had significantly longer overall survival time when compared with those with low P-EZH2-T487 (P=0.032). In multivariate analysis, P-EZH2-T487 was an independent prognostic marker for overall survival in pancreatic cancer (HR=0.358, P=0.001). These results suggest that P-EZH2-T487 is a prognostic indicator for clinical outcome in pancreatic cancer patients.

Citation Format: Yongkun Wei, Shih-Shin Chang, Huamin Wang, Weiya Xia, Mien-Chie Hung. Phosphorylation of EZH2 at Thr487 is a marker for good prognosis in pancreatic cancer patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5351. doi:10.1158/1538-7445.AM2013-5351

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5352: Dysregulation of the repressive H3K27 trimethylation mark in head and neck squamous cell carcinoma contributes to dysregulated squamous differentiation.
Authors: Gannon Orla M. , Merida de Long Lilia , Endo Munoz Liliana , Hazar Rethinam Mehlika , Saunders Nicholas
Published: August 2014

Head and neck squamous cell carcinoma (HNSCC) is one of the most prevalent cancers diagnosed worldwide and is associated with a 5-year survival rate of 55%. EZH2, a component of the polycomb repressor complex 2, trimethylates H3K27 (H3K27me3); which has been shown to drive squamous differentiation in normal keratinocytes. This study determined whether inhibition of EZH2-mediated epigenetic silencing could induce differentiation or provide therapeutic benefit in HNSCC. We determined the effects of inhibiting EZH2, by either RNA interference or pharmacologically, on HNSCC growth, viability and differentiation in vitro. Xenografts of HNSCC cell lines were used to assess efficacy of 3-Deazaneplanocin A (DZNep), an inhibitor of H3K27 trimethylation, in vivo.

EZH2 was highly expressed in HNSCC cell lines in vitro and tissue microarray analysis revealed high expression in (n= 59) in situ relative to normal oral epithelium (n=12). Inhibition of EZH2 with siRNA could induce expression of differentiation genes in differentiation-refractory squamous cell carcinoma cell lines. Differentiation-refractory HNSCC cell lines displayed persistent H3K27me3 on the promoters of differentiation genes. DZNep caused cancer-cell specific apoptosis in addition to a profound reduction in colony forming efficiency and induction of some squamous differentiation genes. Furthermore, in vivo, DZNep attenuated tumour growth in two different xenograft models, caused intra-tumour inhibition of EZH2 and induction of differentiation genes in situ.

Collectively, these data suggest that aberrant differentiation in HNSCC may be attributed to epigenetic dysregulation and suggests that inhibition of PRC2-mediated gene repression may represent a potential therapeutic target.

Citation Format: Orla M. Gannon, Lilia Merida de Long, Liliana Endo Munoz, Mehlika Hazar Rethinam, Nicholas Saunders. Dysregulation of the repressive H3K27 trimethylation mark in head and neck squamous cell carcinoma contributes to dysregulated squamous differentiation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5352. doi:10.1158/1538-7445.AM2013-5352