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miR-765 regulates hepatocellular carcinoma cell proliferation through targeting INPP4B
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Abstract:
Objective:To explore the regulating effect of miR-765 on inositol polyphosphate 4-phosphatase type Ⅱ（INPP4B） and its influence on the proliferation of hepatocellular carcinoma cells (HCC). Methods:Quantitative Real-time PCR was used to detect the expression of miR-765 in 8 pairs of HCC tissues and adjacent tissues as well as its expression in 8 sets of hepatocellular carcinoma cell lines. Methyl thiazol tetrazolium (MTT) assay was used to detect the influence of miR-765 over-expression on proliferation of HCC cell line. Molecular biology technique was used to construct 3′-UTR reporter gene of INPP4B, and to analyze the regulation effect of miR-765 on INPP4B; Western blotting was used to examine the effect of miR-765 over-expression/silencing on INPP4B protein expression; Colony formation assay together with Western blotting were applied to investigate the specific inhibition of INPP4B on the proliferation of HCC cell lines. Results: miR-765 was highly expressed in HCC tissues and HCC cells (P<0.05). miR-765-mimics transfection significantly increased the proliferation capacity of HCC cell line (\[3.78±1.25\] vs \[2.06±0.47\], P<0.05) . miR-765 could significantly down-regulate the luciferase expression of pGL3-INPP4B-3′UTR reporter gene (\[042±0.01\] vs \[101±0.01\], P<0.05) and also up-regulate the expression of INPP4B (\[0.92±0.04\] vs \[0.42±0.02\], \[0.62±003\]，P<0.05). Specific inhibition of INPP4B significantly promoted the proliferation of HCC cells (\[238.0±173\] vs \[66.33±5.04\], P<0.05). Conclusion: miR-765 promotes the proliferation of HCC cells by down-regulating INPP4B expression.
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Project Supported:
Project supported by the Science and Technology Foundation of Jiangxi Province（No.20151BBG70085），and the Education Foundation of Jiangxi Province（No.GJJ14681）
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