摘要
目的研究人巨细胞病毒(HCMV)感染对宫颈癌细胞Siha发生上皮-间质转化的作用及意义。方法实验组用HCMV AD169(MOI=5)和5ng/ml TGF-β1作用宫颈癌Siha细胞,同时将仅TGF-β1作用的Siha细胞作为对照组,分别在0、24、48、72h观察细胞形态的变化,应用RT-PCR检测E-钙粘蛋白和波形蛋白的转录水平,采用Western blot检测E-钙粘蛋白、波形蛋白和MMP-2的表达水平。结果实验组细胞于48h发生细胞形态学的变化,早于对照组的72h;实验组上皮细胞标记物E-钙粘蛋白的转录和表达水平较对照组显著下调(P<0.01),间质细胞标记物波形蛋白的转录和表达较对照组显著上调(P<0.01)且表达提前,MMP-2的表达较对照组显著上调(P<0.01)且表达提前。结论 HCMV感染可促进宫颈癌细胞Siha发生化上皮-间质转化。
Objective To explore the effects of human cytomegalovirus infection on the epithelial-mesenchymal transition in SiHa cervical carcinoma cells. Method The proteins E-cadherin and vimentin and the molecule matrix metalloprotein-2are associated with the epithelial-mesenchymal transition.Changes in expression of E-cadherin,vimentin,and matrix metalloprotein-2were detected.Cells of the human cervical carcinoma cell line SiHa were cultured.Cells in the experimental group were treated with 5ng/ml TGF-β1and HCMV AD169（MOI=5）while the control group was stimulated with TGF-β1without HCMV infection.Changes in cell morphology were observed at 0h,24 h,48h,and 72 h.The levels of transcription of E-cadherin and vimentin were detected with RT-PCR,and the levels of expression of E-cadherin,vimentin,and matrix metalloprotein-2were detected with Western blotting. Results Results indicated that morphological changes occurred in the experimental group at 48 h,which was earlier than changes occurring at 72 hin the control group.The level of transcription of the epithelial cell marker E-cadherin in the experimental group was significantly downregulated（t=7.61,P〈0.01）while the level of transcription of the mesenchymal cell marker vimentin in the experimental group was up-regulated（F=11.5,t=5.23,P〈0.01）.The level of E-cadherin expression was significantly downregulated in the experimental group（t=10.27,P〈0.01）while the level of vimentin expression was up-regulated（F=18.7,t=6.93,P〈0.01）and vimentin was expressed earlier than in the control group.Similar trends were noted regarding the level of matrix metalloprotein-2expression（F=21.0,t=3.91,P〈0.01）. Conclusion Human cytomegalovirus infection may promote the epithelial-mesenchymal transition in SiHa cervical carcinoma cells.
出处
《中国病原生物学杂志》
CSCD
北大核心
2015年第12期1086-1090,1096,共6页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.81070501)
