miR-217靶向E2F3抑制非小细胞肺癌细胞增殖并诱导凋亡

Effect of miR-217 targeting E2F3 on inhibiting the proliferation and apoptosis of non-small cell lung cancer cells

摘要:

目的： 研究miR-217对非小细胞肺癌细胞增殖和凋亡的作用及机制。方法： RT-qPCR检测非小细胞肺癌组织和细胞中miR-217与E2F3 mRNA的表达，利用Lipofectamine 2000将miR-NC、miR-217 mimic、miR-217 inhibitor、pc-NC、pc-E2F3质粒分别或联合转染进入非小细胞肺癌SK-MES-1或A549细胞，克隆形成实验检测细胞增殖，流式细胞术检测细胞凋亡，双荧光素酶报告检测靶向关系，Western blot检测E2F3蛋白相对表达水平。结果： miR-217在非小细胞肺癌组织和细胞中均明显下调（P<0.01）。与Control组相比，miR-217 mimic组非小细胞肺癌SK-MES-1或A549细胞克隆数目明显减少，细胞凋亡率升高（P<0.01），miR-217 inhibitor组非小细胞肺癌SK-MES-1或A549细胞克隆数目明显增加，细胞凋亡率降低（P<0.01）。miR-217靶向下调E2F3。共转染pc-E2F3后逆转miR-217对非小细胞肺癌SK-MES-1或A549细胞增殖和凋亡的作用。结论： miR-217过表达靶向E2F3抑制非小细胞肺癌SK-MES-1或A549细胞增殖并诱导其凋亡。

Abstract:

Objective: To investigate the effect and mechanism of miR-217 on proliferation and apoptosis of non-small cell lung cancer cells. Methods: The expression levels of miR-217 and E2F3 mRNA in non-small cell lung cancer tissues and cells were detected by RT-qPCR.miR-NC, miR-217 mimic, miR-217 inhibitor, pc-NC, pc-E2F3 plasmids were transfected into non-small cell lung cancer SK-MES-1 or A549 cells, respectively or in combination, using Lipofectamine 2000. Cell proliferation was detected by clone formation experiments, apoptosis was detected by flow cytometry, targeting relationship was detected by dual luciferase report, and the relative expression level of E2F3 protein was detected by Western blot. Results: miR-217 was significantly down-regulated in non-small cell lung cancer tissues and cells (P<0.01). Compared with Control group, the number of cloned cells of SK-MES-1 or A549 cells in miR-217 mimic group significantly decreased, and the apoptosis rate increased significantly (P<0.01); the number of cloned cells of SK-MES-1 or A549 cells of non-small cell lung cancer in miR-217 inhibitor group significantly increased and the apoptosis rate decreased (P<0.01).miR-217 targeted down-regulation of E2F3. Co-transfection of pc-E2F3 reversed the effects of miR-217 on proliferation and apoptosis of non-small cell lung cancer SK-MES-1 or A549 cells. Conclusion: Overexpression of miR-217 inhibits the proliferation and induces apoptosis of non-small cell lung cancer SK-MES-1 or A549 cells by targeting E2F3.