miRNA-186调控CDK6和IL-10表达抑制乳腺癌MCF-7细胞生长与转移机制的研究

The inhibition of the cell growth and metastasis of breast cancer cell strain MCF-7 by miRNA-186 targeting CDK6 and IL-10

目的探讨miR-186介导的CDK6和IL-10表达对乳腺癌MCF-7细胞增殖、凋亡及侵袭的影响。方法用已构建的miR-186 mimic质粒转染乳腺癌MCF-7细胞,荧光显微镜下观察转染效率,Western blot技术检测重组质粒对CDK6和IL-10表达的影响,光学显微镜下观察细胞生长群落,分别用MTT法、流式细胞术和Transwell小室法测定重组质粒转染对细胞增殖、凋亡周期及侵袭能力的影响。结果 miR-186转染效率良好,与non-targeting组比较,在转染miR-186后,可明显抑制乳腺癌MCF-7细胞的增殖(P<0.01),细胞生长群落分析结果显示,miR-186组细胞群落数目明显低于non-targeting组细胞(P<0.01),细胞周期结果显示,miR-186组的G0/G1和S期百分比明显的低于non-targeting组(P<0.05),G2/M期百分比明显的高于non-targeting组(P<0.05),Transwell检测结果显示,miR-186组乳腺癌MCF-7细胞侵袭细胞数量(81.00±2.00)个明显少于non-targeting组(125.00±3.00)个,miR-186转染后,CDK6和IL-10在miR-186组乳腺癌MCF-7细胞中的表达率明显低于non-targeting组。结论 miR-186可下调乳腺癌MCF-7细胞中CDK6和IL-10的表达,可有效抑制乳腺癌MCF-7细胞的增殖、侵袭,miR-186可能成为一个潜在的乳腺癌基因治疗靶标。

This study was designed to investigate the effects of miR-186-mediated CDK6 and Il-10 expression on the proliferation,apoptosis and invasion of breast cancer MCF-7 cells.miR-186 mimic plasmid was adopted to transfect MCF-7 cells,and the transfection efficiency was observed with fluorescence microscope.Western blot was used to detect recombinant plasmid expression of CDK6 and IL-10,while optical microscope was used to observe th cell growth.The effects of recombinant plasmid transfection on the proliferation,apoptosis,and invasive ability of MCF-7 cells were evaluated by MTT method,flow cytometry and Transwell Chambers method.The data showed that miR-186 transfection could significantly inhibit the proliferation of breast cancer MCF-7 cells(P<0.01).The results of cell growth community analysis showed that the number of cell communities in the miR-186 group was significantly lower than that in the non-targeting group(P<0.01).The percentage of G2/M stage cells was significantly higher than that of the non-targeting group(P<0.05).Transwell test results showed that the number of breast cancer MCF-7 cells in the miR-186 group(81.00±2.00)was significantly lower than that in the non-targeting group(125.00±3.00).After transfection with miR-186,the expression rate of CDK6 and IL-10 in breast cancer MCF-7 cells of the miR-186 group was significantly lower than that the non-targeting group.In conclusion,miR-186 can down-regulate the expression of CDK6 and IL-10 in breast cancer MCF-7 cells and effectively inhibit the proliferation and invasion of breast cancer MCF-7 cells,thus miR-186 may become a potential target for breast cancer gene therapy.