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目的生存素(survivin)短发夹RNA(shRNA)重组质粒转染B细胞淋巴瘤Raji细胞,体内、外观察对该基因表达的干扰作用及抑制细胞增殖的效果。方法实验分为空白对照(D)组、A组、B组和C组,分别用生存素干扰重组质粒A、B和阴性对照重组质粒转染Raji细胞,经G418筛选获得各重组质粒稳定转染细胞,RT-PCR比较各组生存素mRNA表达差异,FCM测定比较各组生存素蛋白的表达,通过细胞计数比较各组细胞增殖能力的差异。结果成功筛选出稳定转染的Raji细胞。与空白对照组相比,质粒A和质粒B对生存素mRNA的抑制率分别为70.01%和46.46%(P<0.05),对蛋白表达抑制率分别为64.15%和41.16%(P<0.05),两干扰组稳定转染细胞的增殖能力明显下降;质粒A对生存素基因的干扰作用较质粒B明显,抑制瘤细胞增殖的效果也更为明显。结论生存素干扰重组质粒A和B能干扰Raji细胞生存素基因表达,抑制B细胞淋巴瘤Raji细胞增殖。
Objective To transfect the survivin short hairpin RNA (shRNA) recombinant plasmid into Raji cells of B cell lymphoma, and to observe its effect on the expression of this gene in vitro and in vivo and its effect of inhibiting cell proliferation. Methods The experiment was divided into blank control group (D), group A, group B and group C, Raji cells were transfected with survivin plasmid A, B and negative control recombinant plasmids respectively, and the recombinant plasmids were stably transfected by G418 The difference of survivin mRNA expression in each group was compared by RT-PCR. The expression of survivin in each group was compared by FCM. The cell proliferation was compared by cell counting. Results Stably transfected Raji cells were successfully screened. Compared with the blank control group, the inhibitory rates of survivin mRNA of plasmid A and plasmid B were 70.01% and 46.46% (P <0.05), and the inhibitory rates of protein expression were 64.15% and 41.16% (P <0.05) Two interference group stably transfected cells significantly decreased proliferation; plasmid A on the survivin gene interference plasmid plasmid B was significantly inhibited the effect of tumor cell proliferation is more apparent. Conclusion Survivin interference with recombinant plasmids A and B can interfere with Raji cell survivin gene expression, inhibit the proliferation of B cell lymphoma Raji cells.