高糖环境下PLGF基因沉默对人绒毛膜外滋养细胞增殖、迁移及侵袭及Nrf2/HO-1信号通路的影响

Effects of PLGF gene silencing on proliferation, migration, and invasion of human extravillous cytotrophoblasts and Nrf2/HO-1 signaling pathway in high glucose environment

目的探究高糖环境下胎盘生长因子(PLGF)基因沉默对人绒毛膜外滋养细胞(EVCT)的影响。方法体外培养HTR-8/SVneo细胞,将细胞分为对照组(不做处理)、空载组(空载质粒转染)及PLGF沉默组(PLGF双切口酶质粒转染)。CCK-8法、划痕实验及Transwell实验检测各组细胞增殖、迁移、侵袭能力;q RT-PCR及Western blot检测各组细胞PLGF、核转录因子红系2相关因子2(Nrf2)、血红素氧合酶1(HO-1)mRNA及蛋白表达情况。结果对照组与空载组细胞增殖抑制率、划痕愈合率、侵袭细胞数比较,差异无统计学意义(P>0.05);与对照组比较,PLGF沉默组细胞增殖抑制率升高,划痕愈合率、侵袭细胞数降低(P<0.05)。对照组与空载组PLGF、Nrf2及HO-1 m RNA及蛋白表达量比较,差异无统计学意义(P>0.05);与对照组比较,PLGF沉默组PLGF、Nrf2及HO-1m RNA及蛋白表达量降低(P<0.05)。结论高糖环境下PLGF基因沉默能够抑制EVCT增殖、迁移及侵袭,其作用机制可能与抑制Nrf2/HO-1信号通路活性有关。

Objective To investigate the effect of placental growth factor(PLGF)gene silencing on human extravillous cytotrophoblasts(EVCT)in high glucose environment.Methods HTR-8/SVneo cells were cultured in vitro,and the cells were divided into control group(no treatment),empty group(empty plasmid transfection)and PLGF silencing group(PLGF double-incision enzyme plasmid transfection).The cell proliferation,migration,and invasion ability of each group were detected by CCK-8 method,scratch test,and Transwell test;the cell mRNA and protein expressions of PLGF,nuclear factor-erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in each group were detected by qRT-PCR and Western blot.Results There were no significant differences in cell proliferation inhibition rate,scratch healing rate,and cell invasion number between control group and empty group(P>0.05);compared with control group,the cell proliferation inhibition rate was increased,the scratch healing rate and the number of invasive cells were decreased in PLGF silencing group(P<0.05).The mRNA and protein expressions of PLGF,Nrf2,and HO-1 were not significantly different between control group and empty group(P>0.05);compared with control group,the mRNA and protein expressions of PLGF,Nrf2,and HO-1 in PLGF silencing group were decreased(P<0.05).Conclusion PLGF gene silencing can inhibit the proliferation,migration,and invasion of EVCT in high glucose environment,and its mechanism may be related to inhibiting the activity of Nrf2/HO-1 signaling pathway.