丙型肝炎病毒核心蛋白不同区段在HEK293T细胞内的定位

Subcellular localization of different truncated forms of hepatitis C virus core protein in HEK293T cells

目的 :为进一步探讨HCV核心蛋白致癌机制 ,观察了HCV核心蛋白不同区段在HEK2 93T细胞内的定位情况。方法 :构建增强型绿色荧光蛋白 (EGFP)和不同长度核心蛋白区段融合表达的pEGFP_C1系列重组载体 ,对HEK2 93T细胞进行瞬时转染 ,并在激光扫描共聚焦显微镜下观察核心蛋白不同区段在细胞内的定位。结果 :全长核心蛋白定位于细胞质 ;核心蛋白 1～ 5 9aa区段完全定位于细胞核 ;5 0～ 14 0aa区段和 1～ 14 0aa区段在细胞核和细胞质中均存在 ;130～ 191aa区段完全存在于细胞质中。结论 :核心蛋白不同区段在细胞内的定位不同 ,将导致其参与细胞调节的途径和功能的不同。

Objective:In order to study the mechanism of carcinogenesis of hepatocellular carcinoma (HCC) caused by hepatitis C virus, the subcellular localization of different truncated forms of HCV core protein was observed.Methods:HEK293T cells were transfected by the pEGFP-C1 recombinant plasmids which expressed fusion protein of enhanced green fluorescence protein (EGFP) and core proteins. The localization of different truncated forms of core protein in HEK293T cells was analyzed by the laser scanning confocal microscope (LSCM). Results:The innate core protein (1-191 aa) located in cytoplasm, while the localization of truncated form of core protein (1-59 aa) was changed from the cytoplasm to nucleus. The truncated forms 50-140 aa and 1-140 aa of core protein were observed in both cytoplasm and nucleus. The last part of core protein (130-191 aa) was completely located in cytoplasm.Conclusion: The localization of difference innate core proteins is different, that must have effects on their functions involved in cellular regulation and pathogenesis.