丙泊酚介导miR-637/STAT3轴调控肝癌细胞生长的相关机制研究

Relavent mechanism of propofol on regulating the growth of hepatocellular carcinoma cells by mediating MiR-637/STAT3 axis

目的:探究丙泊酚介导微小RNA-637/信号转导及转录活化因子3(miR-637/STAT3)调控肝癌细胞的生长及其机制。方法:正常培养人肝癌细胞株HepG2,随机分为对照组和丙泊酚低、中、高剂量(10、25、50μg·ml^(-1))处理组,通过细胞计数试剂盒-8(CCK-8)实验检测细胞增殖能力,流式细胞仪检测细胞凋亡,转移小室实验检测细胞侵袭能力,划痕实验检测细胞迁移能力,蛋白质印迹法检测细胞中切割后天冬氨酸特异性半胱氨酸蛋白水解酶3(cleaved caspase-3)、B淋巴细胞肿瘤-2(Bcl-2)、Bcl-2相关蛋白X(Bax)、基质金属蛋白酶(MMP)-2、MMP-9、STAT3及磷酸化STAT3(p-STAT3)蛋白表达情况,实时荧光定量聚合酶链反应(qRT-PCR)检测miR-637表达水平。结果:与对照组比较,丙泊酚处理组增殖抑制率、凋亡率、cleaved caspase-3及miR-637表达升高,侵袭细胞数、迁移率、Bcl-2/Bax、MMP-2、MMP-9及p-STAT3/STAT3表达降低(P<0.05),具有剂量依赖性。结论:丙泊酚可介导miR-637/STAT3轴,抑制肝癌细胞的增殖、侵袭与迁移,并诱导其凋亡。

Objective:To explore the mechanism of propofol on regulating the growth of hepatocellular carcinoma(HCC)cells by mediating microRNA-637/signal transducer and activator of transcription 3(miR-637/STAT3).Methods:The human HCC cell line HepG2 was cultured normally and randomly divided into control group,low-dose,medium-dose and high-dose propofol(10,25,50μg·ml^(-1))treatment groups.The proliferation,apoptosis,invasion and migration ability of the cells were respectively detected by CCK-8,flow cytometry,Transwell assay and scratch assay.The expressions of cleaved cysteinyl aspartate specific protease 3(cleaved caspase-3),B-cell lymphoma-2(Bcl-2),Bcl-associated X(Bax),matrix metalloproteinases(MMP)-2,MMP-9,STAT3 and phosphorylated STAT3(p-STAT3)proteins in cells were detected by Western blot.The expression level of miR-637 was determined by real-time quantitative polymerase chain reaction(qRT-PCR).Results:Compared with the control group,the rate of proliferation inhibition and cell apoptosis,expressions of cleaved caspase-3 and miR-637 were increased,while the number of invasion cells,migration rate,expressions of Bcl-2/Bax,MMP-2,MMP-9 and p-STAT3/STAT3 were decreased in the propofol treatment groups(P<0.05)in a dose-dependent manner.Conclusion:Propofol can inhibit the proliferation,invasion and migration of HCC cells,and induce their apoptosis by mediating miR-637/STAT3 axis.