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目的研究miR-143对宫颈癌HeLa细胞增殖和凋亡的影响,探讨miR-143对宫颈癌HeLa细胞系Bcl-2蛋白及mRNA表达的调节作用。方法将miR-143,anti-miR-143以及microRNA空对照高表达质粒用脂质体转染进宫颈癌He-La细胞中,MTT法检测宫颈癌HeLa细胞增殖的变化,His-tone/DNA ELISA检测细胞凋亡;Western blot和RT-PCR检测Bcl-2蛋白和mRNA的表达情况;双荧光素酶报告基因法检测miR-143的靶位点。结果 MiR-143抑制HeLa细胞的增殖,促进细胞凋亡,而anti-miR-143促进HeLa细胞增殖,抑制细胞凋亡;高表达的miR-143抑制Bcl-2蛋白的表达,anti-miR-143增加Bcl-2蛋白的表达量,而Bcl-2 mRNA的变化很小;miR-143抑制Bcl-2 3’端的荧光素酶的活性,突变miR-143与Bcl-2的预测结合位点后,荧光素酶的活性恢复。结论 MiR-143至少部分通过打靶Bcl-2抑制宫颈癌HeLa细胞增殖,并且促进HeLa细胞凋亡。
Objective To investigate the effect of miR-143 on the proliferation and apoptosis of cervical cancer HeLa cells and to explore the role of miR-143 in regulating the expression of Bcl-2 protein and mRNA in cervical cancer HeLa cell line. METHODS: The miR-143, anti-miR-143 and microRNA empty control high expression plasmids were transfected into cervical cancer He-La cells by lipofectamine 2000. The proliferation of HeLa cells was detected by MTT assay. His-tone / DNA ELISA The expression of Bcl-2 protein and mRNA was detected by Western blot and RT-PCR. The target site of miR-143 was detected by dual luciferase reporter assay. Results MiR-143 could inhibit the proliferation of HeLa cells and promote the apoptosis of HeLa cells. However, anti-miR-143 could promote the proliferation of HeLa cells and inhibit the apoptosis of HeLa cells. MiR-143 inhibited the expression of Bcl- Bcl-2 mRNA and Bcl-2 mRNA were significantly increased. MiR-143 inhibited luciferase activity at the 3 ’end of Bcl-2 and mutated the predicted binding sites of miR-143 and Bcl- Luciferase activity restored. Conclusions MiR-143 inhibits the proliferation of cervical cancer HeLa cells at least partly by targeting Bcl-2, and promotes the apoptosis of HeLa cells.