抑制Rac1的活性对人肝癌细胞生长及侵袭能力的影响

目的 探讨抑制人肝癌细胞HepG2中Rac1的活性对其生长及侵袭能力的影响.方法 应用逆转录病毒载体,将Rac1基因的阴性突变体Rac1N17导入HepG2细胞,经嘌呤霉素筛选后获得表达Rac1N17的阳性细胞克隆Rac1N17-HepG2,荧光显微镜下观察绿色荧光蛋白的表达,四甲基偶氮唑(MTT)法检测细胞增殖抑制并绘制细胞生长曲线,细胞基质黏附实验检测对细胞的黏附能力的影响,Transwell法检测细胞侵袭能力的改变.结果 荧光镜下可见绿色荧光蛋白主要分布于HepG2细胞核中,在细胞质中也有少量表达,证实了病毒栽体质粒转染成功.MTT法表明Rac1的活性抑制后,HepG2细胞的增殖能力无明显变化(P>0.05);细胞基质黏附实验显示HepG2细胞与基质的黏附能力随着时间的延长而逐渐减弱(P<0.05);Transwell侵袭实验结果 显示Rac1的活性抑制后,Rac1抑制组穿过PET滤膜的HepG2细胞数明显少于对照组(P<0.05),表明其侵袭能力明显受抑制.结论 Rac1基因在肝癌细胞的黏附、侵袭及过程中发挥重要的作用,有望成为肝细胞癌基因治疗新的靶点.

Effect of the activity of Rac1 gene restrained in HCC on its proliferation and invasive ability

Objective To explore the effect of the activity of Rac1 gene restrained in HepG2 cells on its proliferation and invasive ability. Methods Dominant negative Rac1N17 was transfected into HepG2 cells with a retroviral vector pBABEGFP, which signed green fluorescent protein, HepG2 cells which expressed Rac1N17 were screen out by puromycin, then fluorescent microscope was used to observed the expression of green flu,"orescent protein, MTT, Cell-Matrix adhesive assay and transwell cell assay were used to test the...