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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 miR-30a in human breast cancer cell lines and its effect on the proliferation, migration and invasion of breast cancer cell lines and to find the target genes. Methods: The expression of miR-30a in human breast cancer cell lines (MCF-7, MDA-MB-231, SK-BR3, MDA-MB-468, MDA-MB-453, T47D) and human breast Expression in normal epithelial cells (HBL-100). The breast cancer cell lines MCF-7-miR-30a and MDA-MB-231-miR-30a overexpressing miR-30a were constructed and the proliferation and migration of breast cancer cells were detected by CCK8 and transwell assay respectively And the impact of invasion. The target protein expression was detected by Western blot and its inhibition was verified by Luciferase dual luciferase reporter system. Results: Compared with normal breast epithelial cells, miR-30a showed low expression in human breast cancer cell lines (P <0.05). The overexpression of miR-30a in MCF-7 and MDA-MB-231 breast cancer cell lines inhibited the proliferation, migration and invasion of breast cancer cell lines to varying degrees (P <0.05). Western blotting and dual luciferase reporter assay both showed that forkhead box protein A1 (FOXA1) is a downstream target of miR-30a, and miR-30a can inhibit its expression. Conclusion: miR-30a is overexpressed in human breast cancer cell lines. Overexpression of miR-30a can inhibit the proliferation, migration and invasion of breast cancer cell lines to varying degrees. FOXA1 is a downstream target gene of miR-30a.