沉默T-cadherin对ox-LDL诱导人绒毛膜滋养层细胞HTR-8/SVneo生物学行为的影响

Effects of T-cadherin silencing on ox-LDL induced biological behavior of human chorionic trophoblast cells HTR-8/SVneo

目的 探讨沉默T-钙黏蛋白(T-cadherin)对氧化低密度脂蛋白(ox-LDL)诱导的人绒毛膜滋养层细胞HTR-8/SVneo生物学行为的影响。方法 将人绒毛膜滋养层细胞HTR-8/SVneo分为空白对照组、ox-LDL组、Tcadherin小干扰RNA(siRNA)阴性对照组、T-cadherin siRNA组。各组细胞转染后,实时荧光定量PCR检测各组Tcadherin mRNA水平;MTT法检测HTR-8/SVneo细胞增殖情况;平板克隆形成实验检测细胞克隆形成情况;流式细胞仪检测细胞凋亡情况;Transwell小室实验、划痕实验分别检测细胞侵袭、迁移能力;蛋白免疫印迹实验检测细胞胱天蛋白酶-3(Caspase-3)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和基质金属蛋白酶-2(MMP-2)、MMP-9蛋白表达水平。结果 与空白对照组相比,ox-LDL组和T-cadherin siRNA阴性对照组T-cadherin mRNA、细胞增殖抑制率、凋亡率、Caspase-3、Bax蛋白表达水平升高,克隆形成率、侵袭细胞数、划痕愈合率、Bcl-2、MMP-2、MMP-9蛋白表达水平降低(P<0.05);与ox-LDL组和T-cadherin siRNA阴性对照组相比,T-cadherin siRNA组T-cadherin mRNA、细胞增殖抑制率、凋亡率、Caspase-3、Bax蛋白表达水平降低,克隆形成率、侵袭细胞数、划痕愈合率、Bcl-2、MMP-2、MMP-9蛋白表达水平升高(P<0.05)。结论 沉默T-cadherin可抑制ox-LDL诱导的HTR-8/SVneo细胞异常凋亡,并促进细胞的增殖、侵袭、迁移能力。

Objective To investigate the effect of silencing T-cadherin on oxidized low density lipoprotein(ox-LDL)induced biological behavior of human chorionic trophoblast HTR-8/SVneo cells.Methods Human chorionic trophoblast cells HTR-8/SVneo were divided into the blank control group,the ox-LDL group,the T-cadherin small interfering RNA(siRNA) negative control group and the T-cadherin siRNA group.After each group was treated accordingly,the expression levels of T-cadherin messenger RNA(mRNA) in HTR-8/SVneo cells were detected by real-time fluorescence quantitative PCR in the four groups.The proliferation of cells in each group was detected by tetramethylazolate method.The clone formation of each group was detected by plate clone formation experiment.The apoptosis of each group was detected by flow cytometry.Transwell chamber test and scratch test were used to detect the invasion and migration ability of cells.The protein expression levels of Caspase-3,Bcl-2 related X protein(Bax),B cell lymphoma-2(Bcl-2) and matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9) of cells in each group were detected by Western blot assay.Results Compared with the blank control group,expression levels of T-cadherin mRNA,proliferation inhibition rate,apoptosis rate,Caspase-3 and Bax protein were increased significantly in the ox-LDL group and the T-cadherin siRNA negative control group,and expression levels of clone formation rate,number of invasive cells,scratch healing rate,Bel-2,MMP-2 and MMP-9 protein were decreased significantly(P <0.05).Compared with the ox-LDL group and the T-cadherin siRNA negative control group,expression levels of T-cadherin mRNA,proliferation inhibition rate,apoptosis rate,Caspase-3 and Bax protein were decreased significantly in the T-cadherin siRNA group,and expression levels of clone formation rate,number of invasive cells,scratch healing rate,Bcl-2,MMP-2 and MMP-9 protein were increased significantly(P<0.05).Conclusion Silencing T-cadherin can inhibit ox-LDL induced abnormal apoptosis of HTR-8/SVneo cells,and promote cell proliferation,invasion and migration.