Chinese Clinical Oncology

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Effects of miR-138 on cell proliferation and apoptosis of bladder cancer via targeting SIRT1

HUANG Zhuoya,MIAO Weixian,WANG Xiaobing,ZHU Shuling,WU Tongtong   

  1. Department of Pathology,Huizhou Central Peoples Hospital,Huizhou 516001,China
  • Received:2017-01-13 Revised:2017-03-09 Online:2017-04-30 Published:2017-04-30

Abstract: Objective To investigate the expression of microRNA-138(miR-138)in bladder cancer T24 cells and to study its effect on proliferation and apoptosis of bladder cancer cells as well as its possible target genes. Methods Real-time quantitative PCR(QPCR) was used to detect the expression of miR-138 in bladder cancer cell line T24 and normal bladder epithelial cells SV-HUC-1. T24 cells were divided into normal control group,miR-138 negative control group and miR-138 transfection group, which were transfected with none, negative control fragment and miR-138 mimics. MTT assay and flow cytometry were performed to determine the effect of miR-138 on cell proliferation and apoptosis. Western blotting was used to detect silent mating type information regulation 2 homolog 1(SIRT1)protein levels in each group. Dual luciferase reporter assay was used to confirm the target relationship between miR-138 and SIRT1. Results The level of miR-138 in bladder cancer T24 cells was 0.57±0.19,lower than 1.00±0.26 of normal bladder epithelial cells,and the difference was statistically significant(P<0.05). The miR-138 level of miR-138 transfection group was 2.59±0.67,higher than 1.00±0.36 of normal control group and 1.08±0.49 of miR-138 negative control group(P<0.05). The proliferation rate of miR138 transfection group was lower than that in miR-138 negative control group and normal control group with statistical significance(P<0.05). Fortyeight hours after transfection,the apoptotic rate of miR-138 transfection group was(29.8±1.9)%,higher than(5.8±1.2)% of normal control group and(7.7±0.9)% of miR138 negative control group(P<0.05). The relative expression of miR138 in miR-138 transfection group was 0.59±0.22,lower than 1.00±0.35 of normal control group and 1.20±0.42 of miR-138 negative control group(P<0.05). The double luciferase reporter assay further confirmed that SIRT1 was a direct target of miR138. Conclusion MiR-138 is decreased in bladder cancer cells,and the over-expression of miR-138 may inhibit the proliferation and promote apoptosis of bladder cancer cells by targeting SIRT1.

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