bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1047: Effect of 1,25-Dihydroxyvitamin D in regulating glutamic-oxaloacetic transaminase 1 (GOT1) and redox balance during breast cancer progression
Authors: Zhou Xuanzhu , Wilmanski Tomasz , Teegarden Dorothy
Published: July 2016

Breast cancer is the second most common cancer among women in the US. The active form of vitamin D, 1α,25 dihydroxyvitamin D (1,25(OH)2D), is proposed to inhibit cellular processes that may prevent breast cancer. Possible pro-oxidative effects of 1,25(OH)2D in breast cancer cells which may selectively promote cell death have been proposed. Glutamic-oxaloacetic transaminase 1 (GOT1) is an essential transaminase in glutamine metabolism. GOT1 converts aspartate into oxaloacetate in the cytoplasm, which is then subsequently converted to malate and pyruvate. NADPH is produced during the last step of this process and increased NADPH/NADP+ ratio has the potential to maintain cellular redox balance. Therefore, it is of interest to study 1,25(OH)2D regulation of GOT1 during cancer progression to understand the role of vitamin D in regulating oxidative stress as a mechanism to prevent breast cancer. We employed untransformed MCF10A human breast epithelial cells (MCF10A), as well as Harvey ras-oncogene or ErbB2 (HER-2/neu) oncogene transformed cells (MCF10A-ras and MCF10A-ErbB2) as cell models for early cancer progression. Results showed that 1,25(OH)2D significantly decreased GOT1 mRNA expression at 48 hours in MCF10A-ras cells and MCF10A-ErbB2 cells by 35 ± 8.8% and 49 ± 10%, respectively, and GOT1 protein expression at 72 hours by 51 ± 4.9% and 51 ± 16%, respectively. The susceptibility of MCF10A-ras cells to hydrogen peroxide induced cell death was significantly increased by 1,25(OH)2D, as determined by measuring viable cell number using MTT assay. We further showed that 96-hour 1,25(OH)2D treatment significantly reduced cellular NADPH/NADP+ ratio by 30 ± 2.8% and GSH/GSSG ratio by 34 ± 2.7% in MCF10A-ras cells. Collectively, these results support that 1,25(OH)2D inhibits GOT1 expression and has pro-oxidative effects in oncogene transformed MCF10A cell lines, which may contribute to its role in preventing breast cancer development.

Citation Format: Xuanzhu Zhou, Tomasz Wilmanski, Dorothy Teegarden. Effect of 1,25-Dihydroxyvitamin D in regulating glutamic-oxaloacetic transaminase 1 (GOT1) and redox balance during breast cancer progression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1047.
Abstract 1057: Bioluminescent assays for investigating cellular energy metabolism
Authors: Leippe Donna M. , Sobol Mary , Valley Mike , Karassina Natasha , Duellman Sarah , Vidugiriene Jolanta , Cali Jim
Published: July 2016

Cellular energy metabolism is recognized to have a central role in many cell processes and is an increasingly important area of study in many research fields such as cancer, immunology, obesity, diabetes and neurology. To facilitate these studies, we have taken advantage of key features of bioluminescence, such as increased sensitivity and wide assay windows, to develop assays for measuring energy metabolites. These assays are easy-to-use plate-based assays with streamlined protocols that require minimal cell sample processing. The assays can be used to measure consumption of major fuel sources such as glucose and glutamine from the cell medium, as well as lactate and glutamate secretion. The sensitivity (1-5pmol/sample), broad linear range (0.1-100 uM) and wide dynamic range (maximum signal-to-background > 100 fold) allow all four metabolites to be measured from the same sample of medium and over time, with earlier detection from fewer cells compared to other methods. The assays are also sensitive enough to detect intracellular levels of glutamine, glutamate and lactate and changes in those levels. Additional assays have been developed to measure glucose uptake and NAD/NADH levels in cells. Analysis of these metabolites provides information regarding the energetic state of the cell. In this poster we provide examples demonstrating the utility of these assays to detect a glycolytic shift in cancer cells, monitor cell growth and differentiation with associated changes in glucose uptake and glycolysis, and evaluate insulin sensitivity in primary adipocytes.

Citation Format: Donna M. Leippe, Mary Sobol, Mike Valley, Natasha Karassina, Sarah Duellman, Jolanta Vidugiriene, Jim Cali. Bioluminescent assays for investigating cellular energy metabolism. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1057.
Abstract 1060: Targeting serine hydroxymethyltransferase 2 (SHMT2) suppresses hepatocarcinoma
Authors: Woo Chern Chiuh , Chen Way Cherng , Philip Lee. Teck Hock
Published: July 2016

Glycolytic intermediate 3-phosphoglycerate branched from glycolysis pathway to fuel serine-glycine biosynthesis pathway. Several studies have shown that glycine consumption is strongly correlated to the proliferation rate of cancer cells. Therefore targeting serine-glycine biosynthesis pathway is a promising strategy due to the fact that many of its genes are up-regulated in cancer cells. These genes include PHGDH, PSAT1 and GLDC. Here, we demonstrate that targeting serine hydroxymethyltransferase 2 (SHMT2), also a member of serine-glycine biosynthesis pathway, is able to suppress hepatocarcinoma both in vitro and in vivo. SHMT2 simultaneously converts serine to glycine and tetrahydrofolate to 5,10-methylenetetrahydrofolate. We found that this gene is up-regulated in Hep3B, HepG2 and Huh-7 liver cancer cells versus THLE2 normal liver cells. Knockdown of SHMT2 effectively suppressed Huh-7 cell growth and tumorigenicity. No tumor was formed after inoculation of SHMT2-knockdown Huh-7 cells in nude mice as compared to 100% tumor formation with control Huh-7 cells. The data from tetracycline-inducible SHMT2-knockdown xenograft mouse model showed that SHMT2 inhibition could reduce tumor growth and tumor incidence as well as delayed tumor onset. We also found that SHMT2-knockdown Huh-7 cells were more vulnerable to the cytotoxicity of doxorubicin, a conventional chemotherapeutic agent, as compared to the control Huh-7 cells. On the other hand, overexpression of SHMT2, however, could not transform THLE2 cells into malignancy suggesting that SHMT2 might not a key driver in liver tumorigenesis. SHMT2 activity is correlated to cancer phenotype as shown by the NMR data via metabolic tracing with 13C-glycine. Supplementation of glycine, rather than 5,10-methylenetetrahydrofolate, could reverse the reduced tumorigenicity caused by SHMT2-knockdown. Taken together, our data support that SHMT2 is a potential target in the treatment of hepatocarcinoma.

Citation Format: Chern Chiuh Woo, Way Cherng Chen, Teck Hock Philip Lee. Targeting serine hydroxymethyltransferase 2 (SHMT2) suppresses hepatocarcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1060.
Abstract 1085: Serum miR-30e and miR-223 are promising non-invasive biomarkers for hepatocellular carcinoma
Authors: Ray Ratna B. , Bhattacharya Sourav , Steele Robert , Di Bisceglie Adrian M. , chakraborty Sounak
Published: July 2016

Hepatocellular carcinoma (HCC) is the world's third most common cancer and causes 600,000 deaths annually. Early detection is the key for improving the survival of HCC patients. Current diagnostic markers, such as alpha fetoprotein or fibroscan have their own limitations. Investigating serum microRNA as biomarkers for early diagnosis of cancer has become an important tool in clinical research. In this study, we tested the hypothesis that serum microRNAs (miRNAs) can be used as a potential biomarker for HCC. We analyzed serum miRNA using reverse transcriptase-quantitative PCR (qRT-PCR) from serum samples. Our study revealed that two miRNAs, miR-30e and miR-223, were expressed at significantly lower levels (P < 0.003) in the sera of HCC patients as compared to healthy volunteers. Further, expression of these miRNA was compared between sera from chronic liver disease (CLD) and HCC patients. miR-30e and miR-223 expression was significantly lower in HCC sera as compared to sera from CLD patients. Both miRNA expression levels were lower in HCC liver biopsy specimens as compared to normal liver RNA. Taken together, our results indicated that serum miR-30e and miR-223 are useful biomarkers of HCC irrespective of etiology.

Citation Format: Ratna B. Ray, Sourav Bhattacharya, Robert Steele, Adrian M. Di Bisceglie, Sounak chakraborty. Serum miR-30e and miR-223 are promising non-invasive biomarkers for hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1085.
Abstract 1095: S100A4 is post-transcriptionally inhibited by miR-505-5p and miR-520c-3p in colorectal cancer
Authors: Mudduluru Giridhar , Ilm Katharina , Fuchs Steffen , Stein Ulrike
Published: July 2016

The calcium binding protein S100A4 induces epithelial mesenchymal transition (EMT), migration, invasion, angiogenesis and metastasis. Its induced expression is reported in several cancer types and correlates with poor prognosis. S100A4 is transcriptionally regulated by Sp1, AP1 and CCAAT/enhancer binding protein β transcription factors and its regulation epigenetically controlled by CpG hyper methylation. However, apart from S100A4 functional and transcriptional regulatory aspects, post-transcriptional regulation is not yet elucidated. In this study, we show that specific microRNAs (miR) target the 3’UTR of S100A4. Luciferase-reporter assays with wild-type and mutated miR-505-5p and miR-520c-3p seed sequences of S100A4-3’UTR confirmed the specificity of targeting. Similarly, overexpression of these miRs reduced the S100A4 and inhibition of these miRs function increased the S100A4 protein significantly. Moreover, co-transfection of either S100A4 expressing vector or siRNA-S100A4 along with the miR-505-5p and miR-520c-3p significantly altered the S100A4 mediated migration and invasion in colorectal cancer (CRC) cell lines (p = 0.05). Both these miRs were down-regulated in CRC cell lines and 5 Aza treatment increased these miRs expression and reduced the S100A4 protein amounts. Moreover, in a panel of CRC resected patient tissues (n = 59), miR-520c-3p significantly down regulated (p = 0.03). miR-520c (located in Chr 19, in a big cluster) upstream region was checked for methylation status using methylation specific PCR and combined bisulfite restriction analysis (COBRA) in CRC cell lines, metastatic and non-metastatic CRC resected tissues. We found the upstream promoter region hyper methylated irrespective of metastasis status of patients and in all CRC cell lines checked. 5 Aza treatment of S100A4 less or no expressing cells line Colo206 and Colo320 showed induced miR-520c expression and reduced S100A4 protein in Colo320. Colo206 cells did not show any alternations in expression of S100A4. Taken together, our findings demonstrate that S100A4 is post-transcriptionally regulated by tumor suppressor genes miR-505c and miR-520c and especially miR-520c expression is epigenetically silenced in CRC.

Citation Format: Giridhar Mudduluru, Katharina Ilm, Steffen Fuchs, Ulrike Stein. S100A4 is post-transcriptionally inhibited by miR-505-5p and miR-520c-3p in colorectal cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1095.
Abstract 1099: Up-regulated miR-96-5p inhibits apoptosis by targeting FOXO1 in hepatocellular carcinoma
Authors: Iwai Naoto , Yasui Kohichiroh , Tomie Akira , Kitaichi Tomoko , Dohi Osamu , Gen Yasuyuki , Itoh Yoshito
Published: July 2016

Hepatocellular carcinoma (HCC) is one of the most common cancers and the third leading cause of cancer death worldwide, especially in Asia and sub-Saharan Africa. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and can act as tumor suppressors or oncogenes. To identify miRNA involved in HCC, we performed a genome-wide miRNA gene expression analysis using the human miRNA microarray (Agilent) in paired tumor and non-tumor tissues from patients with primary HCC. The array-based miRNA expression profiles were validated by quantitative PCR. We identified miR-96-5p as the most significantly upregulated miRNA in HCC tumors. Although miR-96-5p has been recognized as an oncogenic miRNA, its role in the pathogenesis of HCC remains unknown. Our study showed that the inhibition of miR-96-5p reduced proliferation of HCC cells and induced apoptosis. Furthermore, we found that the forkhead box O1 (FOXO1) gene was a target of miR-96-5p. FOXO1, a transcription factor, plays an essential role in cell fate decisions. FOXO1 can induce apoptosis through mitochondria-dependent and mitochondria-independent pathways. Collectively, our results suggested that miR-96-5p may function as an oncogenic miRNA through inhibiting apoptosis by targeting FOXO1 in HCC.

Citation Format: Naoto Iwai, Kohichiroh Yasui, Akira Tomie, Tomoko Kitaichi, Osamu Dohi, Yasuyuki Gen, Yoshito Itoh. Up-regulated miR-96-5p inhibits apoptosis by targeting FOXO1 in hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1099.
Abstract 110: Molecular profiling of Tudor domain-containing proteins identifies PHF20L1 as a candidate oncogene in breast cancer
Authors: Jiang Yuanyuan , Liu Lanxin , Shan Wenqi , Yang Zeng-Quan
Published: July 2016

Tudor domain-containing proteins (TDRDs), which recognize and bind to methyl-lysine/arginine residues on histones and non-histone proteins, play critical roles in regulating chromatin architecture, transcription, genomic stability, and RNA metabolism. Dysregulation of several TDRDs have been observed in various types of cancer. However, neither the genomic landscape nor clinical significance of TDRDs in breast cancer has been explored comprehensively. Here, we performed an integrated genomic and transcriptomic analysis of 41 TDRD genes in breast cancer (TCGA and METABRIC datasets) and identified associations among recurrent copy number alterations, gene expressions, clinicopathological features, and survival of patients. Among seven TDRDs (PHF20L1, ARID4B, SETDB1, LBR, TDRKH, TDRD10, and TDRD5) that had the highest frequency (>10%) of gene amplification, the plant homeodomain finger protein 20-like 1 (PHF20L1) was the most commonly amplified (17.62%) TDRD gene in TCGA breast cancers. Different subtypes of breast cancer had different patterns of copy number and expression for each TDRD. Notably, amplification and overexpression of PHF20L1 were more prevalent in aggressive basal-like and Luminal B subtypes and were significantly associated with shorter survival of breast cancer patients. Furthermore, knockdown of PHF20L1 inhibited cell proliferation in PHF20L1-amplified breast cancer cell lines. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains. Detailed characterization of PHF20L1 in breast cancer revealed that the Tudor domain likely plays a critical role in promoting cancer. Mechanistically, PHF20L1 might participate in regulating DNA methylation by stabilizing DNA methyltransferase 1 (DNMT1) protein in breast cancer. Thus, our results demonstrated the oncogenic potential of PHF20L1 and its association with poor prognostic parameters in breast cancer. Furthermore, our findings provide strong foundations for further research and therapeutic targeting of PHF20L1 or other TDRDs in breast cancer.

Citation Format: Yuanyuan Jiang, Lanxin Liu, Wenqi Shan, Zeng-Quan Yang. Molecular profiling of Tudor domain-containing proteins identifies PHF20L1 as a candidate oncogene in breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 110.
Abstract 1101: The role of miR-601 in prostate cancer progression
Authors: Fleming Jessica L. , Hlavin Bell Erica , Andrews Kathryn , Chakravarti Arnab
Published: July 2016

Prostate cancer (PCa) is the second most common cancer among men worldwide. In order to advance treatment options for these men, it is crucial to understand the molecular underpinning behind this cancer. Previously, our group identified miR-601 as a predictive biomarker in PCa. Few studies to date have functionally validated molecular biomarkers and currently nothing is known about the function of miR-601 in PCa. Based on our previous publication, we hypothesized that miR-601 is playing a role in PCa progression. In order to assess this, in vitro experiments were conducted in three PCa cell lines, DU-145, PC-3, and LNCaP. miR-601 was over-expressed and knocked down in each cell line and assays were performed evaluating cell proliferation, colony formation, and apoptosis. Additionally, to provide more information regarding miR-601-associated pathways, we identified putative gene targets of miR-601 using in silico prediction programs, microrna.org and TargetScan.org and evaluated top gene targets in vitro. The results of our experiments indicate that miR-601 is playing a role of a tumor suppressor. Over-expression of miR-601 in cell lines resulted in a significant reduction in cell viability. This was confirmed both by MTS as well as colony formation assays. We looked into the mechanism behind the reduction in cell viability and found that cells over-expressing miR-601 had reduced levels of full-length caspase-3, signifying that these cells are undergoing apoptosis. Two putative gene targets of miR-601 were identified and investigated in vitro, SIRT1, a histone deacetylase known to be both an oncogene and tumor suppressor, and BCL2L2, an anti-apoptotic gene known to be an oncogene. SIRT1 and BCL2L2 had strong scores on both online prediction programs as likely targets of miR-601. Our in vitro results confirmed this. We saw reduced mRNA and protein expression of these targets in cells over-expressing miR-601. Our data thus far suggest that miR-601 is acting as a tumor suppressor in PCa. Targeted therapies for miR-601 and/or its targets may be promising in the treatment of PCa, however additional work is needed to warrant this.

This work was supported by R01CA108633 (To AC), 1RC2CA148190 (To AC) U10CA180850-01 (To AC), 1R01CA169368 (To AC) from the National Cancer Institute (NCI), Brain Tumor Funders Collaborative Grant (To AC), Ohio State University Comprehensive Cancer Center Award (To AC).

Citation Format: Jessica L. Fleming, Erica Hlavin Bell, Kathryn Andrews, Arnab Chakravarti. The role of miR-601 in prostate cancer progression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1101.
Abstract 1106: Mir-223-5p works as an oncomir in vulvar carcinoma by TP63 regulation
Authors: Rocha Rafael , De Melo Maia Beatriz , Rodrigues Iara S. , Amaral Nayra S. , Ling Hui , Calin George A. , Soares Fernando A.
Published: July 2016

Recently, miR-223-5p was associated with metastasis in HPV negative vulvar samples and in several other tumor types. In the present study, we hypothesized that this microRNA would have a potential impact on in vitro vulvar cancer behavior. We artificially mimicked microRNA expression in vulvar cancer cell line, SW962, derived from lymph node metastasis of vulvar carcinoma, and performed in vitro assays as follows in the result section. Impaired cell proliferation (p<0.01) and migration (p<0.001) were observed when miR-223-5p was overexpressed. In contrast, increased invasive potential of the cells was observed (p<0.004). By Luciferase assay we provide evidence that miR-223-5p targets TP63 through its 3’UTR direct ligation (p<0.05). We also demonstrate that this microRNA is capable to decrease levels of p63 at both mRNA and protein levels (p<0.001, and p<0.0001; respectively). We further demonstrated that low p63 protein expression in samples from patients was correlated with deeper tumor (p<0.0345) invasion and lower overall survival (p<0.0494). Our study highlights the complexity surrounding p63 biological mechanisms and the importance of this protein as a prognostic factor in vulvar cancer. We also point out miR-223-5p overexpression as a putative pathological mechanism of tumor invasion and a promising therapeutic target. Also, it is plausible that the evaluation of p63 expression in vulvar cancer at the biopsy level may bring important contribution on prognostic establishment and in elaborating better surgical approaches for vulvar cancer patients.

Citation Format: Rafael Rocha, Beatriz De Melo Maia, Iara S. Rodrigues, Nayra S. Amaral, Hui Ling, George A. Calin, Fernando A. Soares. Mir-223-5p works as an oncomir in vulvar carcinoma by TP63 regulation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1106.
Abstract 1107: miR-29b acts as an oncogene or tumor suppressor gene depending on the external stimulus
Authors: Vassella Erik , Langsch Stephanie
Published: July 2016

microRNAs (miRNAs), short regulatory sequences at the posttranscriptional level, are important mediators of signaling pathways that act as backups of transcriptional control. To identify miRNAs implicated in epidermal growth factor receptor (EGFR) signaling, transformed bronchial epithelial BEAS-2B cells were retrovirally transduced with KRASG12V and alterations in miRNA expression were assessed by microarray analysis. Here we show that miR-29b is significantly induced by mutant KRAS in bronchial epithelial and non-small cell lung cancer (NSCLC) cell lines as well as in primary NSCLC tissue. In agreement with these results, inhibitors of EGFR and MEK resulted in reduced levels of miR-29b while inhibitors of PI3K had no effect. KRASG12V-transduced BEAS-2B cells were significantly more protected from extrinsic apoptosis than control transduced cells, but co-transduction of cells with KRASG12V and anti-miR-29b constructs sensitized cells to apoptosis indicating that miR-29b is a mediator of KRAS-induced resistance to apoptosis. Protection from extrinsic apoptosis was due to enhanced nuclear factor κB (NF-κB) activity. The ubiquitin-editing enzyme tumor necrosis factor alpha-induced protein 3 (TNFAIP3/A20) is a negative regulator of NF-κB signaling. Enhanced NF-κB activity elicited by miR-29b was due to targeting TNFAIP3/A20. Overexpression of miR-29b-refractory TNFAIP3 restored NF-κB activity as well as extrinsic apoptosis, demonstrating that TNFAIP3 is a relevant target of miR-29b. Interestingly, miR-29b conferred sensitivity to cisplatin-induced intrinsic apoptosis by targeting Mcl-1. Thus, miR-29b tips the balance from extrinsic apoptosis towards intrinsic apoptosis. Our results indicate that miR-29b can act either as an oncogene or tumor suppressor gene depending on the external stimulus.

Citation Format: Erik Vassella, Stephanie Langsch. miR-29b acts as an oncogene or tumor suppressor gene depending on the external stimulus. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1107.
Abstract 1108: MicroRNA-211 enhances the oncogenicity of oral carcinoma through targeting TCF12 and up-regulating FAM213A expression
Authors: Chen Yi-Fen , Chang Kuo-Wei , Lin Shu-Chun
Published: July 2016

To address the oncogenic roles of miR-211, we generated K14-EGFP-miR-211 transgenic mouse lines. Upon the 4-nitroquinoline 1-oxide (4NQO) induction, the transgenic mice exhibited more extensive and severe tumorigenesis in tongue compared to the controls. 4NQO up-regulated miR-211 expression in oral squamous cell carcinoma (OSCC) cells. miR-211 directly targeted transcription factor 12 (TCF12), which mediated suppressor activities and was drastically down-regulated in OSCC tumors. GeneChip analysis, bioinformatic algorithm, reporter assay and ChIP assay pinpointed that Family with Sequence Similarity 213, Member A (FAM213A), an anti-oxidative molecule, was transcriptionally repressed by TCF12. Knockdown of FAM213A decreased oncogenic activity and ALDH1-positive cell population, and increased oxygen stress in OSCC cells. The expression of TCF12 and FAM213A was inversely correlated in head and neck carcinoma samples according to The Cancer Genome Atlas. OSCC patients carrying tumors with high FAM213A expression tended to have worse survival. Furthermore, 4NQO treatment down-regulated TCF12 and up-regulated FAM213A through modulating miR-211. The transgenic mouse model recaptures the molecular and histopathological changes as seen in human OSCC pathogenesis. This study highlights that miR-211 responds to oncogenic stimuli to promote the neoplastic process of OSCC by targeting TCF12 and up-regulating FAM213A.

Citation Format: Yi-Fen Chen, Kuo-Wei Chang, Shu-Chun Lin. MicroRNA-211 enhances the oncogenicity of oral carcinoma through targeting TCF12 and up-regulating FAM213A expression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1108.
Abstract 111: Recurrent mutations of CDKN2A genes characterize worse prognosis of HPV positive and negative oropharyngeal squamous cell carcinoma
Authors: Lee Jongkeun , Hong Dongwan , Lee Eun Kyung , Jung Yuh-Seog , Park Weon Seo
Published: July 2016

Background: Oropharyngeal squamous cell carcinoma (OSCC) exhibits unique clinical and molecular characteristics, depending on its association with high-risk human papillomavirus (HPV) or other carcinogen like smoking1. Considering significant subset of patients with OSCC still exhibits poor response to contemporary treatments2, which frequently cause significant toxicity, a more detailed understanding of these poor responders, especially adopting current precision genomic technology, could provide a clue for developing better treatment.

Methods: In this study, we gathered the somatic point mutations and mRNA expression data of 520 OSCC patients from The Cancer Genome Atlas (TCGA) and then investigated recurrent somatic point mutations. To genes having the recurrent and same type point mutations, in addition, we check out the mRNA expression and analyzed a survival analysis.

Results: We found 7 somatic point mutations of CDKN2A gene, which having recurrent patients and only nonsense SNVs. Out of these mutation, 6 recurrent point mutations of CDKN2A gene have highly expressed on PIK3CA, FGFR2 and FGFR3 genes, as well as showed a significant deterioration on survival outcome.

Conclusion: Recurrent nonsense mutations of CDKN2A mostly (6/7, 85.7%) derived an abnormal regulation of other genes, such as PI3KA, FGFR2, and FGFR3. Moreover, this pattern of mutations significantly characterizes ominous prognosis in HPV positive and negative OSCC. Our result might provide better insights for characterizing the subgroup of OSCC and eventual development of precision strategy for this disease subset.

References

1. Licitra, L. et al. High-risk human papillomavirus affects prognosis in patients with surgically treated oropharyngeal squamous cell carcinoma. J Clin Oncol 24, 5630-6 (2006).

2. Ang, K.K. et al. Human papillomavirus and survival of patients with oropharyngeal cancer. N Engl J Med 363, 24-35 (2010).

Citation Format: Jongkeun Lee, Dongwan Hong, Eun Kyung Lee, Yuh-Seog Jung, Weon Seo Park. Recurrent mutations of CDKN2A genes characterize worse prognosis of HPV positive and negative oropharyngeal squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 111.
Abstract 1110: Inhibition of histone deacetylase induces miR-320-mediated androgen receptor suppression in prostate cancer
Authors: Sato Shinya , Katsushima Keisuke , Shinjo Keiko , Takahashi Satoru , Kondo Yutaka
Published: July 2016

Targeting androgen receptor (AR) by pharmacological intervention is one of the effective approaches for treatment of malignant prostate cancers. Histone deacetylase (HDAC) alters epigenetic status of tumor-associated genes, including microRNAs (miRNAs), and affects the behavior of cancers. Here we examined molecular effects of a HDAC inhibitor, OBP-801, on AR expression and tumor cell growth in prostate cancers. Treatment of OBP-801 efficiently suppressed cell growth of three prostate cancer lines (22Rv1, VCaP, and LNCaP), together with AR downregulation, regardless of their hormone sensitivity. Intriguingly, this effect by OBP-801 is not due to downregulation of transcriptional activity of the AR gene, but due to post-transcriptional regulation, namely miRNA-mediated suppression. Among the upregulated miRNAs after OBP-801 treatment in three prostate cancer cell lines, miR-320a, of which expression was significantly correlated with prognosis of prostate cancers (P = 0.0185), was the most closely associated with AR expression. miR-320a mimic suppressed AR protein expression together with growth suppression, while anti-miR320a significantly abrogated the growth suppression by OBP-801 treatment. Fluorescent in situ hybridization analysis revealed that miR-320a was highly expressed in human normal prostate luminal cells but rarely expressed in prostate cancer cells. In AR-dependent prostate tumorigenic rat model, OBP-801 treatment profoundly increased the miR-320 expression and repressed prostate tumorigenesis. Our data demonstrated that OBP-801 effectively suppressed AR activity via epigenetic upregulation of miR-320a, which resulted in tumor cell growth suppression of prostate cancers. OBP-801 may be a potent drug as an AR targeting therapy in AR positive prostate cancer regardless of androgen dependency.

Citation Format: Shinya Sato, Keisuke Katsushima, Keiko Shinjo, Satoru Takahashi, Yutaka Kondo. Inhibition of histone deacetylase induces miR-320-mediated androgen receptor suppression in prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1110.
Abstract 1112: miR-550a-x fuctions as a tumor suppressor by inhibiting the oncogenic YAP1 in human cancers
Authors: Choe Min Ho , Kim Joon , Han Young-Hoon , Kim Jae-Sung
Published: July 2016

Although Yes-associated protein 1 (YAP1), an essential downstream effector of hippo pathway, is a potent oncogene and amplified in human cancers, the overexpression mechanism is unclear. Herein, this study shows that miR-550a-x directly inhibits the oncogenic YAP1 and functions as a tumor suppressor in various human cancer cells. Ectopic overexpression of miRNA-550a-x suppressed proliferation and survival of various cancer cells via inducing apoptosis. miR-550a-x also had a capacity to suppress cell migration, epithelial-mesenchymal transition and tumor sphere formation. Using bioinformatics and microarray analysis, we found YAP1 as a putative target gene. miR-550a-x decreased the mRNA and protein levels of YAP1 in various cancer cells and significantly repressed YAP1 activity with its downstream target genes including CTGF and CYR61. The expression level of miRNA-550a-x were inversely correlated with the YAP1 expression in normal and colorectal cancer samples. Therefore, our data suggest that miRNA-550X functions as a tumor suppressor by targeting YAP1, which may provide a novel therapeutic option for cancer treatment.

Citation Format: Min Ho Choe, Joon Kim, Young-Hoon Han, Jae-Sung Kim. miR-550a-x fuctions as a tumor suppressor by inhibiting the oncogenic YAP1 in human cancers. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1112.
Abstract 1113: Mir-194-3p suppresses tumorigenic potential of glioblastoma stem cells by targeting the canonical NF-kB pathway
Authors: Singh Rajbir , Palanichamy Kamalakannan , Jacob John R. , Chakravarti Arnab
Published: July 2016

Glioblastoma (GBM) is the most common tumor of the central nervous system, with its deadly nature owing to high rates of invasion and angiogenesis. A key element in its pathogenesis is a small subpopulation of neoplastic cells called glioma stem cells (GSCs) that are responsible for tumor initiation, maintenance and relapse. Approaches to target GSCs have mostly proved futile due to their chemo- and radio-resistance potential. Given that microRNAs carry a potential to affect both normal and stem cells, we employed a microRNA-based targeted approach to identify the subset of microRNAs that could distinctively affect the stemness of GSCs. Our study tests the hypothesis that GSCs will show an altered miRNA expression upon their differentiation, and their profiling may lead to identification of specific microRNAs that regulate the process. Our initial efforts resulted in identification of hsa-miR-194-3p as one of the potential candidates, which was consistently downregulated in multiple GSCs that we investigated. Stable cell lines overexpressing hsa-miR-194-3p showed a decreased proliferation rate and reduced sphere-forming ability. In silico analysis revealed TAB2, an upstream activator of NF-κB pathway to be the potential target of hsa-miR-194-3p, which was confirmed by luciferase assay and knockdown approaches. The regulatory role of hsa-miR-194-3p was substantiated by its ability to inhibit the NF-κB pathway and the corresponding downstream targets including IL-6 and STAT3. In vivo studies revealed a reduced tumor forming ability when the cell lines overexpressing hsa-miR-194-3p were injected intracranially in NOD/SCID mice, compared with the non-target control. Our results indicate that hsa-miR-194-3p blocked the canonical NF-κB signaling and negatively affected tumor formation. We have carried out this study this in a unique model system expecting it to uncover another layer of complexity that may add a new facet to meet the challenge imposed by the cancer stem cell paradigm. In conclusion, we accentuate the anti-tumorigenic role of hsa-miR-194-3p as a connecting link to both stemness and NF-κB signaling.

Funding sources: This work was supported by R01CA108633 (To AC), 1RC2CA148190 (To AC) U10CA180850-01 (To AC), 1R01CA169368 (To AC) from the National Cancer Institute (NCI), Brain Tumor Funders Collaborative Grant (To AC), Ohio State University Comprehensive Cancer Center Award (To AC).

Citation Format: Rajbir Singh, Kamalakannan Palanichamy, John R. Jacob, Arnab Chakravarti. Mir-194-3p suppresses tumorigenic potential of glioblastoma stem cells by targeting the canonical NF-kB pathway. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1113.
Abstract 1119: Tumor suppressing dual-action miRNA: Targeting Warburg effect and androgen receptor function in CRPC
Authors: Ebron Jey Sabith , Singh Jagjit , Sikand Kavleen , Shankar Eswar , Gupta Sanjay , Shukla Girish C.
Published: July 2016

The ability of prostate cancer cells to alter their metabolism in a manner distinct from benign cells and energy reprogramming is a major concern in treatment of castration resistant prostate cancer (CRPC). On the other hand, development of resistance to next-generation drugs directed against the AR signaling axis (Abiraterone acetate and Enzalutamide) confers a grim prognosis. Hence, targeting aerobic glycolysis of tumor cells as well as AR signaling axis in CRPC is highly desirable. Here, we report the role of a dual-action miRNA which targets the Androgen Receptor function as well as action of key factors involved in tumor energy metabolism. In addition, the miRNA also targets the key anti-apoptotic factors and enhance the apoptosis in cultured cells. Furthermore, together with Enzalutamide, the miRNA shows a potent synergistic anti-tumor activity in 22RV1 mouse xenografts. We will present the detailed results showing the therapeutic potential of dual-action miRNA in the treatment of CRPC.

Citation Format: Jey Sabith Ebron, Jagjit Singh, Kavleen Sikand, Eswar Shankar, Sanjay Gupta, Girish C. Shukla. Tumor suppressing dual-action miRNA: Targeting Warburg effect and androgen receptor function in CRPC. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1119.
Abstract 1120: Differential expression of human cytomegalovirus microRNA in triple-negative breast cancer tumors
Authors: Hudson Tia , Harrison Scott , K C Dukka , Prins Jan , Muganda Perpetua M.
Published: July 2016

Triple-negative breast cancer (TNBC) accounts for 15-20% of the breast cancer cases.

These tumors are heterogeneous and aggressive, fail to respond to targeted therapy, have poor prognosis, and a high risk of relapse. The molecular basis of this breast cancer subtype is currently unknown. Viruses, such as human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), and human papillomavirus (HPV), are major suspects in the etiology of breast cancer. Since miRNAs have been implicated in the pathogenesis of breast cancer, including triple-negative breast cancer, we hypothesized that viral miRNAs may play a role in this aggressive disease. The objective of this project, therefore, was to determine the identity, prevalence, sequence variation, and differential expression of viral miRNAs in TNBC tumors as compared to control samples. We conducted a comprehensive profiling of viral miRNAs in 48 TNBC tumors as compared to 15 control normal breast tissues, utilizing deep sequencing analysis software and publically available deep sequencing data. Five novel putative HCMV miRNAs were found to be differentially expressed in TNBC tumors as compared to controls. Two of the putative miRNAs were differentially expressed in 60-79% of the TNBC tumors as compared to 0% of normal controls. Two additional putative miRNAs were expressed in 67-88% of TNBC tumors as compared to 25-31% of normal controls. One putative miRNA was expressed in 67% of TNBC tumors as compared to 88% of normal samples, while 1 putative miRNA in addition to HCMV-US25-1-3p displayed no significant differences. These putative HCMV miRNAs localize within genomic regions of HCMV that encode UL56 DNA packaging terminase subunit 2, single-stranded DNA-binding protein, noncoding RNA4.9, and enveloped glycoprotein M. Ongoing work has so far computationally validated one of these miRNAs as a novel HCMV miRNA. This is the first report on the differential expression of HCMV miRNAs in TNBC tumors. Since TNBC tumors are extremely heterogeneous, it is intriguing that 60-80% of TNBC tumors specifically express the same HCMV miRNA. Our findings suggest that these differentially expressed HCMV miRNAs may potentially play a role in the pathogenesis of TNBC.

Citation Format: Tia Hudson, Scott Harrison, Dukka K C, Jan Prins, Perpetua M. Muganda. Differential expression of human cytomegalovirus microRNA in triple-negative breast cancer tumors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1120.
Abstract 1127: KIT/D816V induces SRC-mediated tyrosine phosphorylation of MITF and altered transcription program in melanoma
Authors: Phung Bengt , Kazi Julhash U. , Lundby Alicia , Bergsteinsdottir Kristin , Sun Jianmin , Goding Colin R. , Jönsson Göran , Olsen Jesper V. , Steingrímsson Eiríkur , Rönnstrand Lars
Published: July 2016

The oncogenic D816V mutation of the KIT receptor is well characterized in systemic mastocytosis and acute myeloid leukemia. Although KITD816V has been found in melanoma, its function and involvement in this malignancy is not understood. Here we show that KITD816V induces tyrosine phosphorylation of the microphthalmia-associated transcription factor (MITF) through a triple protein complex formation between KIT, MITF and SRC family kinases. In turn, phosphorylated MITF activates target genes that are involved in melanoma proliferation, cell cycle progression, suppression of senescence, survival and invasion. By blocking the triple protein complex formation, thus preventing MITF phosphorylation, the cells become hypersensitive to SRC inhibitors. We have therefore delineated the mechanism behind the oncogenic effects of KITD816V in melanoma and provide a rationale for testing SRC family kinase inhibitors to treat KITD816V-transformed melanomas.

Citation Format: Bengt Phung, Julhash U. Kazi, Alicia Lundby, Kristin Bergsteinsdottir, Jianmin Sun, Colin R. Goding, Göran Jönsson, Jesper V. Olsen, Eiríkur Steingrímsson, Lars Rönnstrand. KIT/D816V induces SRC-mediated tyrosine phosphorylation of MITF and altered transcription program in melanoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1127.
Abstract 1131: Uncovering oncogenic KRAS allele specific phenotypes using an isogenic MEF cell line system
Authors: Fer Nicole , Smith Brian , Garvey Leslie , Burgan William , Powell Katie , Sharma Kanika , Waters Andrew , Wu Xiaolin , Soppet Dan , Stephens Robert , Nissley Dwight , Holderfield Matthew , Bagni Rachel K.
Published: July 2016

The RAS Initiative at the Frederick National Laboratory has generated an isogenic mouse embryonic fibroblast (MEFs) cell line panel containing single transgene alleles using the RAS-less MEF system developed by Matthias Drosten and Mariano Barbacid (CNIO). These MEFs are H, N and K-RAS null, growth is dependent on exogenous RAS or MAPK pathway activation. The panel includes: KRAS 4A WT, KRAS 4B WT, KRAS 4B G12C, G12D, G12V, G13D, Q61L, Q61R, HRAS WT, NRAS WT or BRAF V600E. The workflow to generate the lines requires cre-lox removal of endogenous KRAS and the cells arrest in the G1 phase of the cycle. Proliferation is resumed through the delivery of the transgene to the cells using lentiviral transduction. Clonal cell lines are derived from the initial pools and are thoroughly characterized including confirmation of endogenous KRAS gene removal, identification of the transgene insertion site(s), calculation of proliferation rates and doubling times, analysis of signaling pathways, response to tool compounds and exome sequenced to exclude lines with mutations in onco-relevant genes. This panel is a unique resource that can be used to ask allele and isoform specific questions (gene expression profiles, GTP-loading, etc.) and screen for RAS inhibitors as well as determine novel compound specificity. This panel is available to the academic RAS research community.

Citation Format: Nicole Fer, Brian Smith, Leslie Garvey, William Burgan, Katie Powell, Kanika Sharma, Andrew Waters, Xiaolin Wu, Dan Soppet, Robert Stephens, Dwight Nissley, Matthew Holderfield, Rachel K. Bagni. Uncovering oncogenic KRAS allele specific phenotypes using an isogenic MEF cell line system. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1131.
Abstract 114: Oncogenomic analysis of Merkel cell carcinoma
Authors: Gelb Tara , Daily Kenneth , Coxon Amy , Brownell Isaac
Published: July 2016

We used an integrative oncogenomic approach to investigate the molecular etiology of Merkel Cell Carcinoma (MCC) in an effort to identify novel therapeutic targets and biomarkers for this highly aggressive neuroendocrine skin tumor. Because MCC is rare and poorly understood, there are currently no effective treatment options for patients with advanced disease. Approximately 80% of MCC tumors have Merkel Cell polyomavirus (MCV) DNA integrated into the host genome, and viral oncogenes are thought to drive carcinogenesis. Moreover it has been reported that MCV-negative MCC samples have an increased rate of mutations relative to MCV-positive tumors. Taken together, this suggests that MCV-positive and negative MCCs are driven by different mechanisms. We used whole exome sequencing and bioinformatic analyses to identify somatically dysregulated genes and pathways in MCC tumor samples. Additionally, we used a comparative genomic hybridization array to identify genomic amplifications and deletions. Whole transcriptome profiling was achieved using microarrays and RNA-sequencing. As the same tumor samples were used in each of these platforms, we were able to systematically correlate sequence variants with copy number changes and gene expression to identify high priority targets for further study. Interestingly, despite differences in driver mutations, we found relatively few differences in the global gene expression of MCV-positive and MCV-negative MCC tumors. Overall, our integrative oncogenomic approach has identified novel therapeutic targets for the treatment of MCC that are currently undergoing functional validation and preclinical testing.

Citation Format: Tara Gelb, Kenneth Daily, Amy Coxon, Isaac Brownell. Oncogenomic analysis of Merkel cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 114.