微小RNA-182靶向调控第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信号通路对肝癌干细胞生物学行为的影响及其机制

目的:观察微小RNA(miR)-182靶向调控第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)信号通路对肝癌干细胞(LCSCs)生物学行为的影响，并探讨其作用机制。方法:采用免疫磁珠干细胞分选系统从人肝癌细胞(HepG2)中分选出LCSCs，然后分为空白对...

Effect of microRNA-182 on the biological behaviors of liver cancer stem cells by targeting the phosphatase and tensin homolog deleted on chromosome ten/phosphoinositide 3-kinase/protein kinase B signaling pathway and underlying mechanism

Objective To observe the effect of microRNA(miR)-182 on the biological behavior of liver cancer stem cells(LCSCs)by targeting the phosphatase and tensin homolog deleted on chromosome ten/phosphoinositide 3-kinase/protein kinase B(PTEN/PI3K/Akt)signaling pathway and explore its mechanism.Methods The LCSCs were separated from HepG2 cells by immunomagnetic bead stem cell sorting system,and then randomly divided into the blank control group,the negative control group and the miR-182 mimics group.The expression level of miR-182 mRNA and the biological behavior changes of all groups after transfection were detected.At the same time,the relative protein expression levels of PTEN/PI3K/Akt signaling pathway were detected.One-way ANOVA was used for comparison of measurement data between multiple groups,LSD-t test was used for pair comparison between groups,andχ2 test was used for comparison of enumeration data.Results The miR-182 mRNA expression level in miR-182 mimics group(3.623±0.702)was higher than that in the blank control group(0.985±0.081)and the negative control group(0.967±0.075),and the difference was statistically significant(t=26.239,27.105,P<0.05),and there was no significant difference between the blank control group and the negative control group(t=0.312,P>0.05).The cell proliferation rate[(17.67±3.52)%,(35.67±7.74)%,(61.36±10.26)%],migration[(47.25±3.83)%]and invasion ability[(207±28)]in miR-182 mimics group at 24,48,72 h after transfection were significantly increased as compared with those in the blank control group[(11.91±1.87)%,(23.12±5.31)%,(40.79±8.71)%;(18.21±1.73)%;(128±17)]and the negative control group[(12.03±1.95)%,(23.80±5.45)%,(41.06±9.02)%;(18.37±1.69)%;(126±15)],with the difference being statistically significant(F=12.304,13.173,14.065;t=29.116,29.305;t=17.390,17.214,P<0.05),but the apoptosis rate at 24,48 and 72 h after transfection[(6.36±0.82)%,(12.07±1.46)%,(20.24±2.83)%]was significantly lower than that in the blank control group[(9.79±1.12)%,(22.18±3.27)%,(38.12±4.85)%]and the negative control group[(10.04±1.25)%,(21.73±3.09)%,(37.92±4.77)%](F=11.512,12.091,12.762,P<0.05),and there was no statistically significant difference in biological behaviors between the blank control group and the negative control group(t=0.277,0.294,0.256,0.364,0.302,0.241,0.332,P>0.05).The protein expression level of PTEN in miR-182 mimics group(0.235±0.024)was lower than that in blank control group(0.494±0.053)and negative control group(0.487±0.057),while the protein expression levels of p-Akt and p-PI3K in miR-182 mimics group(0.871±0.110,0.396±0.042)were higher than those in blank control group(0.320±0.036,0.122±0.017)and negative control group(0.325±0.039,0.127±0.019).The difference was statistically significant(t=13.512,13.473;t=21.074,21.023;17.211,17.192,P<0.05).There was no significant difference in the expression levels of Akt and PI3K among the groups(t=0.341,0.230,0.371,0.434,0.442,0.274,P>0.05),and there was no significant difference in the expression levels of PTEN,p-Akt and p-PI3K between the blank control group and the negative control group(t=0.265,0.271,0.375,P>0.05).Conclusion MiR-182 may activate the PI3K/Akt signaling pathway by inhibiting PTEN,thereby enhancing the proliferation,migration and invasion ability of LCSCs and inhibiting apoptosis of LCSCs,and it may serve as a new target for clinical treatment of hepatocellular carcinoma.