miR-124通过调控PI3K/Akt信号通路对人胃癌MGC803细胞增殖与凋亡的影响  

The effect of miR-124 on proliferation and apoptosis of human gastric cancer MGC803 cells by regulating PI3K/Akt signaling pathway

作　　者：罗远[1] 蒋海忠[1] Luo Yuan

机构地区：[1]浙江省宁波市第一医院,315200

出　　处：《浙江临床医学》2020年第10期1407-1410,共4页Zhejiang Clinical Medical Journal

基　　金：浙江省医药卫生科技项目(2019KY578)。

摘　　要：目的探讨miR-124对人胃癌MGC803细胞增殖、凋亡的影响及调控PI3K/Akt信号通路的作用机制。方法培养人胃癌细胞株MGC803,将miR-124模拟物(miR-124mimics组)及miR-124阴性对照(miR-124 NC组)转染至MGC803细胞中,并采用CCK-8法及细胞克隆实验考察转染后MGC803细胞增殖情况,流式细胞术检测转染后细胞凋亡,Western blotting检测PI3K、Akt、p-PI3K和p-Akt蛋白水平,RT-qPCR检测PI3K和Akt mRNA表达。结果转染miR-124后,miR-124mimics组中miR-124水平为(4.15±0.18),显著高于miR-124 NC组(P<0.01)。miR-124mimics组转染48h和72h后的OD450值分别为(0.86±0.08)和(1.08±0.13),均显著低于miR-124 NC组(P<0.05),且其细胞克隆形成数显著降低(P<0.01);转染48h和72h后MGC803细胞凋亡率为(51.80±8.14)%及(70.40±7.14)%,显著高于miR-124 NC组(P<0.01)。同时,miR-124mimics组PI3K/Akt信号通路中关键靶点PI3K、Akt、p-PI3K和p-Akt蛋白水平显著低于miR-124 NC组(P<0.05);miR-124mimics组PI3K/Akt信号通路中PI3K和Akt基因表达水平显著低于miR-124 NC组(P<0.01)。结论miR-124可抑制人胃癌MGC803细胞的增殖,促进细胞凋亡,其可能通过下调PI3K/Akt信号通路发挥作用。Objective To investigate the effect of miR-124 on proliferation and apoptosis of human gastric cancer MGC803 cells and the mechanism of regulating PI3K/AKT signaling pathway.Methods Human gastric cancer cell line MGC803 was cultured.The miR-124mimics(miR-124 mimics group)and miR-124 negative control(miR-124 NC group)were transfected into MGC803 cells.CCK-8 assay and cell clone assay were used to investigate the proliferation of MGC803 cells.The apoptosis of MGC803 cells was detected by using flow cytometry.The protein levels of PI3K,Akt,p-PI3K and p-Akt were detected via western blotting,and mRNA expressions of PI3K and Akt were detected by RT-qPCR.Results The levels of miR-124 in miR-124 mimics group was(4.15±0.18),which was significantly higher than that in miR-124 NC group(P<0.01).The OD450 values of miR-124 mimics group at 48h and 72h after transfection were(0.86±0.08)and(1.08±0.13),respectively,which were sigificantly lower than those in miR-124 NC group(P<0.05),and whose number of cell clone formation was decreased signifcantly(P<0.01).And the apoptosis rate of MGC803 cells in miR-124 mimics group at 48h and 72h after transfection were(51.80±8.14)%and(70.40±7.14)%,respectively,significantly higher than those in miR-124 NC group(P<0.05).At the same time,the levels of PI3K,Akt,p-PI3K and p-Akt proteins from PI3K/Akt signaling pathway in miR-124 mimics group were significantly lower than those in miR-124 NC group(P<0.05).The expression of PI3K and Akt genes from PI3K/Akt signaling pathway in miR-124 mimics group was significantly lower than those in miR-124 NC group(P<0.01).Conclusion The miR-124 can inhibit the proliferation and promote apoptosis of human gastric cancer MGC803 ell,which may play a role by down-regulating PI3K/Akt signaling pathway.

关 键 词：miR-124 人胃癌MGC803细胞 细胞增殖 细胞凋亡 PI3K/AKT信号通路 

分 类 号：R73[医药卫生—肿瘤]