摘要
人巨细胞病毒(Human cytomegalovirus,HCMV)感染能够影响胎盘绒毛外滋养细胞的增殖、侵袭,进而参与病理妊娠的发生,但具体的机制尚未阐明。在真皮成纤维细胞中的研究表明,HCMV能够抑制Wnt/β-catenin通路。因此,为了观察HCMV感染通过Wnt/β-catenin通路抑制滋养细胞增殖及侵袭的作用,本研究培养了人绒毛外滋养细胞株HTR8/SVneo。研究分为对照组、HCMV组、HCMV+LiCl组细胞。对照组细胞用培养液处理,HCMV组细胞用含有3个感染复数(Multiplicity of infection,MOI)HCMV的培养液处理,HCMV+LiCl组细胞用含有3MOI HCMV及50mmol/L Wnt/β-catenin通路激活剂LiCl的培养液处理。48小时后,采用MTS法检测细胞增殖,采用Transwell检测细胞侵袭,采用western blot检测细胞周期蛋白D1(cyclin D1)、基质金属蛋白酶2(Matrix metalloproteinase 2,MMP2)、基质金属蛋白酶9(Matrix metalloproteinase 9,MMP9)、β-连环蛋白(β-catenin)和磷酸型糖原合酶激酶-3β(p-GSK-3β)的表达量。结果显示,与对照组细胞相比,HCMV组细胞的O.D_(490)水平、侵袭数目及cyclinD1、MMP2、MMP9、β-catenin、p-GSK-3β的表达量均降低(P<0.05);与HCMV组细胞比较,HCMV+LiCl组细胞的O.D_(490)水平、侵袭数目及cyclinD1、MMP2、MMP9、β-catenin、p-GSK-3β的表达量均增加(P<0.05)。以上结果表明,HCMV感染抑制人绒毛外滋养细胞的增殖和侵袭,且该作用与抑制Wnt/β-catenin通路有关。本研究阐明了HCMV感染通过抑制Wnt/β-catenin信号通路的激活而抑制滋养细胞增殖及侵袭的分子机制,为最终阐明HCMV先天感染的致病机理积累了有价值的研究资料。
Human cytomegalovirus(HCMV)infection can affect the proliferation and invasion of extravillous trophoblasts in the placenta,and participate in pathological pregnancy,but the exact mechanism is not known.A study on dermal fibroblasts showed that the HCMV can inhibit the Wnt/β⁃catenin pathway.We wished to observe the effects of HCMV infection on inhibition of proliferation and invasion of trophoblasts via the Wnt/β⁃catenin pathway.A human extravillous trophoblast cell line(HTR8/SVneo)was cultured and divided into three groups:control,HCMV,and HCMV+LiCl.Cells in the control group were treated with medium.Cells in the HCMV group were treated with medium containing HCMV at a multiplicity of infection(MoI)of 3.Cells in the HCMV+LiCl group were treated with medium containing HCMV(3 MoI)together with an activator of the Wnt/β⁃catenin pathway:LiCl(50 mmol/L).Then,48 hours later,the proliferation and invasion of trophoblasts as well as expression of cyclin D1,metalloproteinase(MMP)⁃2,MMP⁃9,β⁃catenin and p⁃GSK⁃3βwere measured.Absorbance at 490 nm,the number of invading trophoblasts,as well as expression of cyclin D1,MMP2,MMP9,β⁃catenin and p⁃GSK⁃3βin cells of the HCMV group were decreased significantly(P<0.05)compared with those of the control group.Absorbance at 490 nm,the number of invading trophoblasts,as well as expression of cyclin D1,MMP2,MMP9,β⁃catenin and p⁃GSK⁃3βwere increased in cells of the HCMV+LiCl group(P<0.05)compared with those in cells of the HCMV group.These results showed that HCMV infection inhibited the proliferation and invasion of human extravillous trophoblasts,and that the effect was related to inhibition of the Wnt/β⁃catenin pathway.We elucidated how HCMV infection regulates the proliferation and invasion of trophoblasts by inhibiting activation of the Wnt/β⁃catenin signaling pathway.Our study provides valuable data to aid understanding of the molecular mechanism of HCMV infection.
作者
蒋英彩
韩毓
韦佩佳
杜就旧
韦先梅
陈婵
JIANG Yingcai;HAN Yu;WEI Peijia;DU Jiujiu;WEI Xianmei;CHEN Chan(Haikou Fourth People's Hospital,Haikou 571199,China)
出处
《病毒学报》
CAS
CSCD
北大核心
2021年第4期803-808,共6页
Chinese Journal of Virology
作者简介
蒋英彩(1987-),女,主治医师,从事妇产科病毒感染疾病研究,E⁃mail:jyc2912@163.com;通讯作者:蒋英彩(1987-),女,主治医师,从事妇产科病毒感染疾病研究,E⁃mail:jyc2912@163.com。
