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探讨EZH2对结肠癌细胞增殖调控作用以及具体作用机制。通过对结肠癌细胞系SW480以及HCT116进行EZH2基因沉默,以检测EZH2对结肠癌细胞的增殖调控作用。MTT检法测细胞的增殖,流式细胞仪检测细胞周期及荧光定量PCR检测周期相关基因Cyclin D1、P15、P21以及mi R-608的表达变化。si RNA转染后,结肠癌细胞中EZH2的表达明显下降(p<0.001),细胞增殖受到明显抑制(p<0.05,p<0.01或p<0.001);si RNA组与阴性对照相比,G_1期细胞比例增高,G_2/M、S期细胞相对减少,cyclin D基因表达下调(p<0.01,p<0.05),P15、P21表达上调(p<0.01)。通过荧光定量PCR发现,结肠癌细胞增殖抑制基因mi R-608在EZH2基因沉默组表达量显著上调(p<0.001)。萤光素酶活性测试结果表明,EZH2在SW480和HCT116细胞中直接调控miR-608的基因转录(p<0.001)。此外,miR-608基因沉默阻断siEZH2对结肠癌细胞增殖的调控作用。本研究发现了EZH2对结肠癌细胞增殖的显著促进作用,其具体作用机制在于抑制miR-608基因表达。因此,EZH2可望成为结肠癌潜在的治疗靶标。
To investigate the effect of EZH2 on the proliferation of colon cancer cells and its mechanism of action. EZH2 gene silencing was performed on the colon cancer cell lines SW480 and HCT116 to test the effect of EZH2 on the proliferation of colon cancer cells. Cell proliferation was detected by MTT assay, cell cycle was detected by flow cytometry, and the expression of Cyclin D1, P15, P21 and mi R-608 was detected by real-time PCR. The expression of EZH2 in colon cancer cells was significantly decreased after transfection with si RNA (p <0.001), and the cell proliferation was significantly inhibited (p <0.05, p <0.01 or p <0.001). Compared with the negative control, (P <0.01, p <0.05), while P15 and P21 were up-regulated (p <0.01). The expression of cyclin D mRNA was down-regulated in G 2 / M and S phase. Fluorescent quantitative PCR showed that the expression of mi R-608, a suppressor of proliferation of colon cancer cells, was significantly up-regulated in the EZH2 gene silencing group (p <0.001). Luciferase activity assay results showed that EZH2 directly regulates miR-608 gene transcription (p <0.001) in SW480 and HCT116 cells. In addition, miR-608 gene silencing blocked the regulation of siEZH2 on the proliferation of colon cancer cells. This study found that EZH2 significantly promote the proliferation of colon cancer cells, its specific mechanism of action is to inhibit miR-608 gene expression. Therefore, EZH2 is expected to become a potential therapeutic target for colon cancer.