钙结合蛋白S100A2对肝癌细胞生长增殖的抑制作用

Calcium-binding protein S100A2 in the retardation of growth and proliferation of hepatocellular carcinoma cells

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 study the retardation effect of calcium binding protein S100A2 on the growth and proliferation of hepatocellular carcinoma QGY7701 cells.Methods After the plasmid of EGFP S100A2 had been regrouped and introduced into the hepatocellular carcinoma QGY7701 cells with lipofectin TM , the expression and location of the products were observed by fluorescent microscopy. The cell growth and proliferation were monitored through cell colony formation in vitro and xenografting subcutaneously in the nude mice in vivo. The effect of S100A2 on the QGY7701 cell cycle was examined with flow cytometry.Results The chimera protein of S100A2 and green fluorescent protein were detected and appeared to be localized in the cytoplasm and nucleus, while the green fluorescent protein was found to be localized only in the cytoplasm. The cell colony formation of QGY7701/A2 was significantly reduced as compared with the controls. The xenografted tumor of QGY7701/A2 in the nude mice showed a growth at a considerably slower rate than that of QGY7701/pc and QGY7701 groups. The cell cycle review showed retardation of QGY7701/A2 cells in the G 1 phase and the DNA content was obviously reduced as compared with the controls.Conclusion The exogenous S100A2 is able to check the QGY7701 cell cycle, stop the cell growth and proliferation either in vitro or in vivo in the QGY7701 cells. [