目的探讨肿瘤转移抑制基因Kai1/CD82对人喉癌Hep-2细胞株增殖能力的影响。方法以喉癌Hep-2细胞株为研究对象,将携带有Kai1基因的重组腺病毒(rAd-Kai1)进行扩增、纯化并测定其滴度,用适量rAd-Kai1转染人喉癌Hep-2细胞株,以转染空病毒载体及未转染Kai1的喉癌细胞株为对照,用四甲基偶氮唑蓝(MTT)法测定Kai1对细胞增殖能力的影响,用RT-PCR技术对转染后的细胞进行检测。结果rAd-Kai1经扩增、纯化后,滴度可达6×10^10PFU/ml,当病毒量为30 MOI时,Hep-2细胞的转染率可达90%以上。MTT实验显示转染空腺病毒组与未转染组比较,Hep-2细胞株体外增殖能力无统计学差异,转染Kai1组Hep-2细胞株体外增殖能力明显弱于未转染组(P〈0.05),抑制率可达29%。结论肿瘤转移抑制基因Kai1/CD82对喉癌细胞株体外增殖能力具有抑制作用。
Objective To study the effect of metastasis suppressor gene Kai1/CD82 on the cell proliferation of Hep-2 cell line.Methods Recombinant adenovirus rAd-Kai1 was amplified,purified,and was used to transfect laryngo-carcinoma Hep-2 cell line.Cells transfected with blank vector and untransfected cells served as controls.MTT assay was used to assess the influence of Kai1 gene on proliferation of Hep-2 cells;RT-PCR was used to examine the expression of Kai1 gene in cells of different groups.Results The titer of rAd-Kai1 reached 6×10^10PFU/ml after expansion and purification.When the concentration of the adenovirus was 30 MOI,the transfection rate of Hep-2 cells could be higher than 90%.MTT assay showed that the cell proliferation abilities were similar in the two control groups,and the proliferation ability of cells in the Kai1 transfection group was significantly lower than that in the non-transfected group(P〈0.05),with the inhibitory rate in the Kai1 group being 29%.Conclusion The Kai1/CD82 gene can inhibit the proliferation of Hep-2 cells.