miR-30a对人乳腺癌细胞系增殖、迁移和侵袭能力的影响

Effects of miR-30a on proliferation, migration and invasion of breast cancer cell lines

目的 :探讨miR-30a在人乳腺癌细胞系中的表达,及其对乳腺癌细胞系增殖、迁移和侵袭的影响,并寻找作用靶基因。方法:用q RT-PCR法检测miR-30a在人乳腺癌细胞系(MCF-7、MDA-MB-231、SK-BR3、MDA-MB-468、MDA-MB-453、T47D)和人乳腺正常上皮细胞(HBL-100)中的表达。构建miR-30a过表达的人乳腺癌细胞系MCF-7-miR-30a、MDA-MB-231-miR-30a,分别用CCK8法、transwell法检测过表达miR-30a对于乳腺癌细胞增殖、迁移和侵袭的影响。通过Western印迹法检测靶蛋白的表达,并通过Luciferase双荧光素酶报告系统验证其抑制作用。结果:与乳腺正常上皮细胞相比,miR-30a在人乳腺癌细胞系中均呈低表达(P<0.05)。在MCF-7、MDA-MB-231乳腺癌细胞系中过表达miR-30a后,乳腺癌细胞系的增殖、迁移和侵袭能力均有不同程度的抑制(P<0.05)。Western印迹法和双荧光素酶报告系统检测均提示,叉头盒蛋白A1(forkhead-box A1,FOXA1)为miR-30a的下游靶基因,miR-30a可靶向抑制其表达。结论:miR-30a在人乳腺癌细胞系中低表达,过表达miR-30a可不同程度抑制乳腺癌细胞系的增殖、迁移和侵袭能力。FOXA1是miR-30a的下游靶基因。

Objective To investigate the expression of micro RNA-30a（mi R-30a） in breast cancer cell lines and the role in cell proliferation, migration and invasion, and the potential target gene. Methods The expression levels of mi R-30 a in breast cancer cell lines（MCF-7, MDA-MB-231, SK-BR3, MDA-MB-468, MDA-MB-453, T47D） and normal human breast epithelial cell line（HBL-100） were detected by quantitative real-time PCR. The mi R-30 a overexpressed cell lines MCF-7-mi R-30 a and MDA-MB-231-mi R-30 a were constructed. The effects of mi R-30 a on cell proliferation were determined by CCK8 assay. The effects of mi R-30 a on migration and invasion abilities were determined by transwell assay. The interested target gene forkhead-box A1（FOXA1） was validated by Western blot and dual-luciferase reporter assay system. Results The expression level of mi R-30 a was significantly lower in breast cancer cell lines than normal breast epithelial cell line（ P 〈0.05）. Overexpression of mi R-30 a of MCF-7 and MDA-MB-231 cells decreased the cell proliferation, migration and invasion abilities at different extent（P〈0.05）. Western blot and dual-luciferase reporter assay system suggested that FOXA1 was one potential target gene negatively regulated by mi R-30 a. Conclusions The lower expression level of mi R-30 a is found in breast cancer cell lines than normal breast epithelial cell line. Overexpression of mi R-30 a can inhibit the proliferation, migration and invasion abilities of breast cancer cell lines. FOXA1 might be the potential target gene regulated by mi R-30 a.