摘要
目的探讨丙型肝炎病毒核心蛋白(HCV C)是否通过调控转录因子NFAT1的表达参与肝内胆管癌的发展及恶性生物学行为。方法构建表达HCV C核心蛋白的真核表达质粒pEGFP-N3-HCV C,通过瞬时转染pEGFP-N3-HCV C质粒,RT-PCR检测肝内胆管癌RBE细胞中NFAT1 mRNA的表达情况,Western blot检测NFAT1蛋白的表达情况,MTT法检测细胞增殖,流式细胞术检测细胞周期变化情况。结果转染HCV C后胆管癌细胞NFAT1基因的mRNA及蛋白表达明显上升,差异具有显著性(P<0.05);HCV C能够促进细胞向G2/M期进展及使细胞增殖能力增加。结论 HCV C可上调胆管癌细胞中NFAT1基因的表达,促进细胞周期进展及胆管癌细胞增殖,可能与肝内胆管癌的发展有关。
Objective To explore whether hepatitis C virus core protein （HCV C） regulates the expression of NFAT1 to participate in the progression and malignant biological behavior of intrahepatic cholangiocarcinoma cells. Methods The recombinant plasmid pEGFP-N3-HCV C and the empty vector pEGFP-N3 were cotransfected with enhanced green fluorescent protein （EGFP） into RBE cells using liposome. Real-time PCR and Western blotting were used to examine the expression of NFAT1 mRNA and protein in the transfected RBE cells. MTT assay was used to evaluate the changes in the cell proliferation, and the cell cycle changes were analyzed by flow cytometry. Results HCV C transfection significantly enhanced the expressions of NFAT1 mRNA and protein in RBE cells （P〈0.05） and promoted the progression of cell cycle into GdM phase to accelerate the cell proliferation. Conclusion Transfection with HCV C gene up-regulates NFAT1 expression and promotes the cell cycle progression and proliferation of intrahepatic cholangiocarcinoma cells, suggesting the involvement of HCV C in the progression of intrahepatic cholangiocarcinoma.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2012年第6期789-793,共5页
Journal of Southern Medical University
基金
国家自然科学基金(30872485)~~
