B7-H3基因敲降ES-2细胞系构建及其对细胞增生的影响

目的 构建B7同系物3(B7 homolog 3,B7-H3)敲降的ES-2细胞系并探究B7-H3基因对ES-2细胞系增生的影响。方法 利用慢病毒介导的短发夹RNA(short hairpin RNA,shRNA)干扰技术在ES-2细胞中敲降B7-H3,利用荧光显微观察、实时荧光定量PCR 技术(real-time fluorescent quantitative PCR,qRT-PCR)及流式细胞术验证B7-H3敲降效果,并利用细胞计数试剂盒(cell counting kit-8,CCK-8)检验细胞活性改变。结果 感染不同的shRNA,可呈现不同程度的B7-H3降低,CCK-8检测结果显示,敲除B7-H3后细胞增生速率明显低于对照组,差异有统计学意义(P<0.05)。结论 使用慢病毒介导的shRNA干扰技术能成功构建B7-H3敲降的ES-2细胞系,B7-H3敲降后ES-2细胞增生速率显著降低。

Construction of B7-H3 knockdown ES-2 cell line and its effect on cell proliferation

Objective To construct a B7-H3 knockdown ES-2 cell line and determine the effect of B7-H3 gene on the proliferation of ES-2 cell line. Methods Lentivirus mediated shRNA interference was used to establish ES-2 cell line with B7-H3 knockdown. Fluorescence microscopy,real-time fluorescent quantitative PCR(qRT-PCR) and flow cytometry were used to verify the effect of B7-H3 knockdown, and cell counting kit-8 (CCK-8) was used to test its effect on proliferation. Results The results of CCK-8 showed that the proliferation rate of the experimental group was significantly lower than that of the control group, with statistical significance(P<0.05). Conclusion B7-H3 knockdown ES-2 cell line is successfully constructed, and it is found that B7-H3 knockdown can significantly reduce the proliferation rate of ES-2 cells.