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

Guo Fan; Zhen Huifen; Ma Lijun

摘要

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/PI3 K/Akt)信号通路调控乳腺癌细胞增殖迁移的机制.方法应用LipofectamineTM 3000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 LipofectamineTM3000.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.

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

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