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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: To further investigate the oncogenic mechanism of HCV core protein, the localization of different regions of HCV core protein in HEK2 93T cells was observed. Methods: A recombinant vector pEGFP_C1 was constructed by fusion expression of enhanced green fluorescent protein (EGFP) and core segments of different lengths. HEK2 93T cells were transiently transfected and observed under different laser scanning confocal microscopy Intracellular localization. Results: The full-length core protein localized in the cytoplasm. The core protein 1 ~ 59a region was completely localized in the nucleus. The 50 ~ 140aa region and the 1 ~ 140aa region were present in the nucleus and cytoplasm. The 130-191aa region Exist in the cytoplasm. Conclusion: The different localization of different regions of the core protein in cells will lead to their different ways and functions of participating in cell regulation.