抗Cyclin D1胞内抗体AD5N对宫颈癌HeLa细胞的影响

Effects of Anti-cyclin D1 Intrabody AD5N on HeLa Cervical Cancer Cells

目的:抗Cyclin D1胞内抗体AD5N对宫颈癌细胞增殖的抑制效应分析。方法:将携带抗Cyclin D1胞内抗体AD5N的真核表达载体pcDNA3.1-AD5N,利用脂质体介导法稳定转染宫颈癌HeLa细胞,G418筛选阳性细胞株,进一步通过Westernblot、MTT法及流式细胞术分析该胞内抗体的细胞内表达和抗肿瘤效应。结果:AD5N在稳定筛选的细胞株中有效表达,与空细胞和空质粒转染的细胞株相比,明显抑制HeLa细胞增殖,并诱导细胞凋亡(P<0.01),细胞周期G0-G1期指数增高10.35%,S期指数下降15.77%,凋亡细胞指数上升6.38%。结论:抗Cyclin D1胞内抗体AD5N基因的表达,抑制宫颈癌细胞的增殖,阻滞细胞周期进展,并诱导细胞凋亡。

Objective： To evaluate the effects of anti-cyclin D1 intrabody AD5N on the proliferation of HeLa cervical cancer cells. Methods： The control plasmids （pcDNA3.1） and recombinant plasmids with intrabody （pcD- NA3.1-AD5N） were stably transfected into HeLa cells using lipofectamine 2000. G418 selection was performed to obtain the positive cell clones that were named HeLa/pcDNA3.1 or Hela/pcDNA3.1-AD5N. Western blot, MTT assay and fluorescence-activated cell sorting were conducted to investigate the expression of intrabody in HeLa cells and its anti-tumor effects. Results： AD5N was expressed efficiently in HeLa/pcD- NA3.1-AD5N cells. Compared with the control, the expression of AD5N in HeLa/pcDNA3.1-AD5N cells significantly inhibited proliferation, markedly arrested the cell cycle, and induced apoptosis of HeLa cells （P〈0.01）. The ratio of cells in G0-G1 phase was increased by 10.35%, and the percentage of cells in S phase was decreased by 15.77%. The apoptosis index was increased by 6.38%. Conclusion： The expression of anti-cyclinD1 intrabody AD5N inhibits cell proliferation, arrests the cell cycle, and induces apoptosis of HeLa cervical cancer cells in vitro.