PEG10对人类绒毛滋养层细胞系HTR-8/Svneo细胞增殖和凋亡影响的初步研究  

Preliminary study on effect of PEG10 on proliferation and apoptosis of human chorionic trophoblastic cell line HTR-8/Svneo

作　　者：张云山 董丽娟 许丽华 马天仲 ZHANG Yun-shan;DONG Li-juan;XU Li-hua;MA Tian-zhong(Reproductive Medicine Center,Affiliated Hospital of Guangdong Medical University,Zhanjiang 524001;Health Management Center of Affiliated Hospital of Guangdong Medical University,Zhanjiang 524001)

机构地区：[1]广东医科大学附属医院生殖医学中心,湛江524001 [2]广东医科大学附属医院健康管理中心,湛江524001

出　　处：《生殖医学杂志》2023年第1期98-105,共8页Journal of Reproductive Medicine

基　　金：湛江市财政资金科技专项竞争性分配项目(2017A01031);广东医科大学附属医院博士启动基金(BJ201804);湛江市科技发展专项资金竞争性分配项目(2022A01184)。

摘　　要：目的初步研究父系表达印记基因PEG10低表达对人类绒毛滋养层细胞系HTR-8/Svneo细胞增殖和凋亡的影响。方法构建4组PEG10干扰载体[si-898、si-1854、si-1951及对照(si-NC)],然后转染到HTR-8/Svneo细胞株并进体外培养,si-NC组为对照组。流式细胞术检测HTR-8/Svneo细胞的细胞周期和细胞凋亡,qRT-PCR检测Cytochrome C、Caspase3、Bid、Bak、Bax及Cyclin D1 mRNA的表达情况,MTT法检测细胞增殖活力。结果与对照组比较,转染3组siRNA片段均能显著抑制PEG10基因的表达(P<0.05),其中si-898、si-1854抑制作用更显著(P<0.01)。转染si-898或si-1854的HTR-8/Svneo细胞Cytochrome C、Caspase3、Bid、Bak、Bax mRNA的表达显著升高(P<0.01),而Cyclin D1 mRNA的表达显著降低(P<0.05),并且细胞凋亡率也显著高于对照组(P<0.05)。MTT法检测细胞增殖活力显示,与对照组相比,干扰PEG10基因后细胞在48 h和72 h的增殖活力也显著降低(P<0.05)。流式细胞术检测显示,干扰PEG10基因后细胞周期阻滞在G1期。结论干扰PEG10基因能诱导人类绒毛滋养层细胞系HTR-8/Svneo细胞凋亡,降低细胞增殖活力,阻止细胞周期从G1期到S期的转化进程,导致细胞周期阻滞。Objective:To study the effects of low expression of paternal imprinted gene PEG10 on the proliferation and apoptosis of human chorionic trophoblast cell line HTR-8/Svneo.Methods:Four groups of PEG10 interference vectors,i.e.si-898,si-1854,si-1951and si-NC,were constructed.HTR-8/Svneo cells were transfected and cultured in vitro,and si-NC served as control group.The cell cycle and apoptosis of HTR-8/Svneo cells were detected by flow cytometry.The mRNA expressions of cytochrome C,Caspase 3,Bid,Bak,Bax and cyclin D1 were detected by RT-PCR.The proliferation activity of HTR-8/Svneo cells was detected by MTT assay.Results:Compared with the control group,the siRNA fragments which were transfected into the three groups significantly inhibited the expression of PEG10 gene(P<0.05),especially si-898 and si-1854(P<0.01).The mRNA expressions of cytochrome C,caspase 3,Bid,Bak and Bax in HTR-8/Svneo cells transfected with si-898 or si-1854 were significantly increased(P<0.05),while the mRNA expression of cyclin D1 was significantly decreased(P<0.05),and the apoptosis rate was significantly increased(P<0.05).MTT assay showed that the cell proliferation activity at 48 hours and 72 hours after the interference of PEG10 was also significantly reduced compared with the control group(P<0.05).Flow cytometry showed that the cell cycle was blocked in G1 phase after interfering with PEG10.Conclusions:Interference with PEG10 can induce apoptosis of human chorionic trophoblast cell line HTR-8/Svneo,reduce cell proliferation,prevent the process from G1 phase to S phase of cell cycle and lead to cell cycle arrest.

关 键 词：不明原因复发性流产 PEG10基因 人类绒毛滋养层细胞系HTR-8/Svneo细胞 凋亡 细胞周期 增殖 

分 类 号：R714.2[医药卫生—妇产科学]