微小RNA-29a通过10号染色体缺失的磷酸酶及张力蛋白同源物/磷脂酰肌醇3激酶/蛋白激酶B信号通路调控乳腺癌细胞增殖迁移的机制

The effect of microRNA-29a on the proliferation and migration of breast cancer cell via phosphatase and tensin homologue deleted on chromosome ten/phosphatidylinositol 3 kinase/protein kinase B signal pathway

目的 探讨微小RNA(miRNA,miR)-29a通过第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因/磷脂酰肌醇3激酶/蛋白激酶B(PTEN/PI3K/Akt)信号通路调控乳腺癌细胞增殖迁移的机制。方法 应用Lipofectamine^TM3000将miR-29a inhibitor、miR-29a NC转染到MCF-7乳腺癌细胞,实时定量聚合酶链反应(Real-time PCR)检测miR-29a表达,细胞计数试剂盒(CCK-8)法检测细胞活力,流式细胞术检测细胞凋亡及细胞周期,Transwell法检测细胞迁移能力,Western blot检测B细胞淋巴瘤/白血病-2(bcl-2)、细胞周期蛋白(Cyclin) D1、基质金属蛋白酶(MMP)-2、MMP-9、PTEN、PI3K、磷酸化Akt(p-Akt)蛋白表达。结果 miR-29a inhibitor组(0.53±0.05)miR-29a表达量较miR-29a NC组(2.74±0.27)上调(P<0.05)。miR-29a inhibitor组(0.35±0.02)细胞活力较miR-29a NC组(0.58±0.05)降低(P<0.05)。miR-29a inhibitor组细胞早期凋亡率(15.38±1.54)%、晚期凋亡率(23.69±2.37)%较miR-29a NC组[(2.53±0.23)%、(3.17±0.31)%]提高(P<0.05)。miR-29a inhibitor组细胞周期G1期较miR-29a NC组延长(P<0.05)。miR-29a inhibitor组(42.87±4.88)细胞迁移数目较miR-29a NC组(136.49±10.37)降低(P<0.05)。miR-29a inhibitor组中bcl-2、Cyclin D1、MMP-2、MMP-9、PI3K、p-Akt表达量较miR-29a NC组下调(P<0.05),PTEN表达量较miR-29a NC组上调(P<0.05)。结论 下调miR-29a表达能通过抑制PTEN/PI3K/Akt信号通路抑制MCF-7乳腺癌细胞增殖与迁移。

Objective To explore microRNA (miRNA, miR)-29a on the proliferation and migration of breast cancer cell MCF-7 via phosphatase and tensin homologue deleted on chromosome ten/phosphatidylinositol 3 kinase/protein kinase B (PTEN/PI3K/Akt) signal pathway.Methods miR-29a inhibitor and miR-29a NC were transfected into MCF-7 cell by liposome Lipofectamine^TM3000. The expression of miR-29a was detected by real-time quantitative polymerase chain reaction (Real-time PCR). Cell viability was detected by cell counting kit-8 (CCK-8) assay. Cell apoptotic rate and cell cycle was detected by flow cytometry. Cell migration ability was detected by Transwell method. The expression of B cell lymphoma/leukemia-2 (bcl-2), Cyclin D1, matrix metalloproteinase (MMP)-2, MMP-9, PTEN, PI3K and phosphorylated Akt (p-Akt) was detected by Western blotting.Results The expression of miR-29a in miR-29a inhibitor group (0.53±0.05) was lower than that in miR-29a NC (2.74±0.27) (P<0.05). Cell viability in miR-29a inhibitor group (0.35±0.02) was lower than that in miR-29a NC group (0.58±0.05) (P<0.05). The early apoptosis rate (15.38±1.54)%, late apoptosis rate (23.69±2.37)% in miR-29a inhibitor group were higher than that in miR-29a NC group [ (2.53±0.23)%, (3.17±0.31)%] (P<0.05). The cell cycle G1 phase in miR-29a inhibitor group was longer than that of miR-29a NC group (P<0.05). Cell migration number in miR-29a inhibitor group (42.87±4.88) was lower than that in miR-29a NC group (136.49±10.37) (P<0.05). The expression of bcl-2, Cyclin D1, MMP-2, MMP-9, PI3K and p-Akt in miR-29a inhibitor group was lower than that in miR-29a NC group (P<0.05). The expression of PTEN in miR-29a inhibitor group was higher than that in miR-29a NC group (P<0.05).Conclusion Down-regulation expression of miR-29a could suppress proliferation and migration in breast cancer MCF-7 cell via inhibition of PTEN/PI3K/Akt signal pathway.