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目的 探讨钙结合蛋白S10 0A2对肝癌细胞生长增殖的抑制作用。方法 构建绿色荧光蛋白 S10 0A2融合基因的重组表达载体 ,经脂质体介导转入QGY770 1肝癌细胞中进行表达 ,应用荧光显微镜直接观察融合蛋白的表达和定位 ;通过肿瘤细胞集落形成实验 ,观察外源性S10 0A2表达对体外培养细胞增殖能力的影响 ;以流式细胞仪分析S10 0A2对肿瘤细胞增殖周期的影响 ;通过裸鼠接种实验 ,观察S10 0A2对肿瘤细胞在体内增殖能力的影响。结果 绿色荧光蛋白 S10 0A2融合蛋白表达、定位在胞浆和胞核 ,而单一的绿色荧光蛋白 (载体对照 )则只定位在胞浆中 ;细胞在液体和半固体培养基中培养的细胞集落形成率 ,实验组 (QGY770 1/A2 )明显低于载体对照组 (QGY770 1/pc)和细胞对照组 (QGY770 1) ;细胞周期分析显示 ,实验组细胞有G1和G2 期阻制 ,DNA合成量明显减少 ;实验组裸鼠体内移植瘤重量明显低于两个对照组。结论 外源性S10 0A2可抑制QGY770 1肝癌细胞在体外的集落形成和在裸鼠体内的增殖能力 ,并对细胞周期有阻滞作用。
Objective To investigate the inhibitory effect of calcium binding protein S10 0A2 on the growth and proliferation of hepatoma cells. Methods The recombinant expression vector of green fluorescent protein S10 0A2 fusion gene was constructed and transfected into QGY770 1 hepatocellular carcinoma cells by liposome. The expression and localization of the fusion protein were directly observed by fluorescence microscope. The effect of exogenous S10 0A2 expression on cell proliferation in vitro was studied. The effect of S10 0A2 on the proliferation cycle of tumor cells was analyzed by flow cytometry. The effect of S10 0A2 on the proliferation of tumor cells was observed in nude mice. Results The green fluorescent protein S10 0A2 fusion protein was expressed in the cytoplasm and nucleus, while the single green fluorescent protein (vector control) was localized only in the cytoplasm; the cell colony cultured in liquid and semi-solid medium was formed. Rate, the experimental group (QGY770 1/A2) was significantly lower than the vehicle control group (QGY770 1/pc) and cell control group (QGY770 1); cell cycle analysis showed that the experimental group cells have G1 and G2 phase inhibition, DNA synthesis Significantly reduced; in the experimental group nude mice implanted tumor weight was significantly lower than the two control groups. Conclusion Exogenous S10 0A2 can inhibit colony formation and proliferation of QGY770 1 hepatocarcinoma cells in nude mice, and block the cell cycle.