摘要
目的 检测戊型肝炎病毒ORF3蛋白对人张氏肝细胞（Changliver,CCL13）增殖及细胞周期的影响.方法 将真核表达载体pDsRed-Monomer-N1-ORF3采用LpofectamineTM2000脂质体法转染CCL13细胞;用MTT比色法测定细胞增殖情况;用流式细胞仪碘化丙啶染色法检测细胞周期;并采用Western-Blot检测细胞周期蛋白的表达.结果 转染pDsRed-Monomer-N1-ORF3的CCL13细胞自第3天开始,增殖受抑（P＜0.01）;细胞周期阻滞于G0/G1期;细胞周期蛋白Cyclin Dl及Cyclin E1的表达受抑,Cyclin A2及Cyclin B1蛋白表达量与对照组无差异.结论 戊型肝炎病毒ORF3蛋白可通过抑制细胞周期蛋白Cyclin Dl及Cyclin E1的表达而抑制CCL13细胞的增殖.
Objective To study the effect of hepatitis E virus Open Reading Frame 3 protein on the proliferation and cell cycle of Chang liver cells （CCL13）.Methods The pDsRed-Monomer-N1-ORF3 recombinant plasmid was transfected into CCL13 cells via lipofectamine 2 000 reagent,then assayed cell proliferation by MTT assay;Flow cytometric was used to detect cell cycle after dying with PI;and the cell cycle proteins expression were detected by western-blotting.Results pDsRed-Monomer-N1-ORF3 recombinant plasmid was successfully transfected into CCL13 cells,Expression of ORF3 can inhibit the proliferation activity of CCL13 cells from the 3rd day （P 〈 0.01）,and prevent the cell cycle in G0/G1 phase;Western Blot showed expression of cell cycle proteins cyclin D1 and cyclin E1 were decreased in ORF3 group compared with the control group;However,the expression of cyclin A2 and cyclin B1 had no significantly difference in three groups.Conclusions HEV-ORF3 can inhibits the proliferation activity of CCL13 cells by inhibition expression of cell cycle proteins cyclin D1 and cyclin E1.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2016年第4期373-377,共5页
Chinese Journal of Experimental and Clinical Virology
基金
基金项目：甘肃省青年科技基金计划（145RJYA247）
国家自然科学基金（81260326）
