Abstract 3966: Regulation of Chd5 expression by microRNA-211 and microRNA-130b in colorectal cancer
2011
Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL
PURPOSE: Chromodomain helicase DNA-binding protein 5 (CHD5) is identified as a novel tumor suppressor in particular of colorectal tumorigenesis. There is limited information about the regulation of CHD5 expression. In this study, we investigated the role of microRNAs (miRNAs) in regulation of CHD5 protein expression in colorectal cancer. DESIGN METHODS: MiR-211 and miR-130b as predictive target CHD5 candidate miRNAs were selected through Targetscan software. A high level of CHD5 protein colorectal cancer HCT-116 cell line was used to establish miR-211 and miR-130b stable transfected cell lines (HCT-116miR-211 and HCT-116miR-130b) as test models, following by MTT, colony assay, flow cytometry for cell viability examination. Quantitative real-time PCR and Western blot analysis were also used for measure miR-211, miR-130b and CHD5 expression levels in HCT-116miR-211 and HCT-116miR-130b. CHD5-associated proteins expression levels were assessed as well. RESULTS: MiR-211 and miR-130b were cloned into lentiviral GFP expression miRNA vector, separately. Retrovirus was produced by using lentiphos TMHT packaging system and infected into HCT-116 cell line. The expression levels of miR-211 in HCT-116miR-211 and miR-130b in HCT-116miR-130b were significantly increased (miR-211 increased 16 times, and miR-130b increased 8 times) and at the same time CHD5 expression levels was decreased. Compared to the vector control cell lines, the levels of cell proliferation and colony formation were higher in HCT-116miR-211 and HCT-116miR-130b. In addition, the proportion of cell number in the G0-G1 phase of HCT-116miR-211 and HCT-116miR-130b was lower than those in the control cell lines. CONCLUSION: Up-regulation of miR-211 and miR-130b induced the malignant phenotype of HCT-116 cells via block of CHD5 expression. That implicated the miRNAs might be one of the mechanisms that lead CHD5 down-regulated in colorectal cancer. This work was supported in part by grants P20 CA118770 and U54 CA091431 from National Cancer Institute.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3966. doi:10.1158/1538-7445.AM2011-3966
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