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目的:探讨miR-124是否通过靶向调节TET蛋白家族的表达而抑制结肠癌细胞增殖与侵袭。方法:用双荧光素酶报告基因检测系统分别检测miR-124对TET家族(TET1、TET2、TET3)的3’UTR-荧光素酶活性的影响;用q RT-PCR与Western blot检测miR-124模拟物转染结肠癌HT29细胞后TET家族的m RNA与蛋白表达水平的变化;用MTS和Transwell实验观察HT29细胞转染miR-124模拟物及TET si RNA后增殖和侵袭能力的变化。结果:双荧光素酶报告基因检测结果显示,各TET m RNA的3’UTR均被miR-124特异性结合,其荧光素酶活性被明显抑制(均P<0.05);转染miR-124模拟物后的HT29细胞TET的m RNA与蛋白表达水平明显减低(均P<0.05);HT29细胞转染miR-124模拟物或TET si RNA后,增殖和侵袭能力均明显降低(均P<0.05)。结论:miR-124可能通过直接靶向调控TET基因的表达,而抑制结肠癌细胞增殖和侵袭。
Objective: To investigate whether miR-124 can inhibit the proliferation and invasion of colon cancer cells by targeting the expression of TET protein family. METHODS: The effect of miR-124 on the 3’UTR-luciferase activity of the TET family (TET1, TET2, TET3) was measured using a dual luciferase reporter gene assay system; miR-124 was detected by qRT-PCR and Western blot. MRNA and protein expression levels of the TET family after transfection of the HT29 cells with the mimic were transfected with MTS and Transwell experiments to observe the changes in proliferation and invasiveness of miR-124 mimics and TET si RNA transfected with HT29 cells. Results: The dual luciferase reporter assay results showed that the 3′UTR of all TET m RNAs were specifically bound by miR-124 and the luciferase activity was significantly inhibited (all P<0.05); miR-124 was transfected The mRNA and protein expression levels of TET in HT29 cells after treatment were significantly reduced (all P<0.05). After transfection of miR-124 or TET si RNA in HT29 cells, the proliferation and invasiveness were significantly reduced (all P<0.05). . CONCLUSION: miR-124 may inhibit the proliferation and invasion of colon cancer cells through direct targeted regulation of TET gene expression.