miR-548c-5p对乳腺癌细胞MCF-7的影响  

Effect of miR-548c-5p on biological behaviors of breast cancer cell line MCF-7

作　　者：侯冷晨[1] 孔祥杰[2] 李佳[2] 张俊峰[2] 李晓宇[2] 房林[2] 

机构地区：[1]同济大学附属第十人民医院麻醉科,上海200072 [2]同济大学附属第十人民医院普外科,上海200072

出　　处：《同济大学学报（医学版）》2013年第4期20-25,共6页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金(61272240)

摘　　要：目的研究miR-548c-5p转染乳腺癌MCF-7细胞株后对细胞增殖、迁移及凋亡的影响。方法 CCK-8试剂盒检测不同浓度的miR-548c-5p(50、75、100、125、150 nmol/L)分别在24、48、72 h不同时间点对细胞增殖的影响;miR-548c-5p转染MCF-7乳腺癌细胞株后,荧光定量PCR法检测其转染效率;划痕实验检测细胞迁移能力的变化;流式细胞术分析干扰后细胞周期的变化;荧光显微镜观察转染后乳腺癌细胞凋亡的变化。结果使用RNA干扰技术成功地将miRNA转染入乳腺癌细胞株,使得miR-548c-5p的表达升高。转染后48 hmiR-548c-5p浓度为125 nmol/L时,对乳腺癌细胞的抑制作用最明显。miR-548c-5p表达上调后细胞G_0/G_1明显增多,而S期明显减少(P<0.01),能一定程度上抑制细胞的迁移。荧光显微镜观察发现早期凋亡细胞升高。结论 miR-548c-5p通过干扰细胞周期,增加G_0/G_1期的乳腺癌细胞数,进而抑制细胞增殖。抑制乳腺癌细胞株MCF-7的迁移能力并显著促进其凋亡。miR-548c-5p影响乳腺癌细胞的增殖、迁移和凋亡,有望成为乳腺癌诊疗的靶点之一。Objective To investigate the effects of miR-548c-5p on biological behaviors of MCF-7 breast cancer cell line. Methods The growth inhibition rates of human breast cancer MCF-7 cells were determined by CCK-8 kits after treated with different concentrations of miR-548c-5p （50, 75, 100, 125 and 150 nmol/L） at different time points （24, 48 and 72 h）. MCF-7 breast cancer cells were transfected with miR-548c-5p by Lipofectamine 2000 and the transfection efficiency was measured by QRT-PCR method. Cell invasion ability was assessed by scratch assay, cell cycle changes were analyzed by flow cytometry, cell apoptosis was observed by fluorescence microscopy in breast cancer MCF-7 cells after transfection with miR-548c-5p. Results The expression of miR-548c-5p was significantly enhanced after transfection （P 〈 0. 01 ）. MiR-548c-5p significantly inhibited the proliferation of MCF-7 breast tumor cells at 48 h after transfection with the concentration of 125 nmol/L. Compared with controls, cells in G0/G1 phase increased, while cells in S phase decreased（ P 〈 0.01 ）. The invasion ability of MCF-7 cells was suppressed and the early apoptotic cells were increased after transfection. Conclusion MiR-548c-5p can increase G0/G1 phase, inhibit cell proliferation and invasion and induce apoptosis of human breast cancer MCF-7 cells, indicating that miR-548c-5p may be used as a new target for treatment of breast cancer.

关 键 词：乳腺肿瘤 miR-548c-5p 凋亡 增殖 迁移 

分 类 号：R737.9[医药卫生—肿瘤]