miR-449a过表达抑制人神经母细胞瘤细胞SH-SY5Y增殖并促进其凋亡

目的探讨miR-449a对人神经母细胞瘤细胞系SH-SY5Y的增殖和凋亡的影响。方法用Lipofectamine TM2000将miR-449a模似物或miR-449a对照转染至SH-SY5Y细胞,分为空白、miR-449a模似物和miR-449a对照SHSY5Y细胞组;实时荧光定量PCR(q-PCR)检测各组细胞中miR-449a表达;CCK8法检测细胞增殖;流式细胞仪检测细胞凋亡和周期,Western blot检测c-Myc蛋白和Bax/Bcl-2蛋白表达。结果 miR-449a模似物瞬时转染SH-SY5Y细胞后,miR-449a的表达水平明显高于正常对照组(P0.05);SH-SY5Y细胞增殖能力受到明显抑制(P0.05);凋亡率明显增加(P0.05);c-Myc蛋白表达显著降低(P0.05);细胞促凋亡蛋白Bax表达升高;抗凋亡蛋白Bcl-2表达降低(P0.05)。结论 miR-449a可通过c-Myc影响SH-SY5Y细胞的增殖和周期,通过调节Bax/Bcl-2影响其凋亡。

Over-expressed miR-449a inhibitsproliferation and induces apoptosis of SH-SY5Y cells in vitro

Objective To explore the effects of miR-449a on the proliferation and apoptosis in human neuroblastoma SH-SY5Y cells.Methods Transferred miR-449a mimics SH-SY5Y cells by LipofectamineTM2000 as experimental group.Cell growth was measured by CCK8 assay,the cell cycle and apoptosis were detected by flow cytometry.Relative expression of c-Myc and Bax/Bcl-2 was detected by Western blot.Results Compare to blank and negative control cells,the proliferation of miR-449a mimics SH-SY5Y cells was decreased(P<0.05).Flow cytometry showed that over-expressed miR-449a resulted in an increase of G0/G1 phase and apoptotic cells.The expression of c-Myc showed significant difference (P<0.05),Expression of pro-apoptotic protein Bax was increased,and that of anti-apoptotic protein Bcl-2 was decreased(P<0.05).Conclusions miR-449a over-expression can inhibit cell proliferation by regulating cell cycles and induce apoptosis mainly through regulating Bcl-2 family proteins expression in SH-SY5Y cells.