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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6×1010PFU/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 investigate the effect of tumor metastasis suppressor gene Kai1 / CD82 on the proliferation of human laryngeal carcinoma Hep-2 cell line. Methods Human laryngeal carcinoma Hep-2 cell line was used as the research object. The rAd-Kai1 carrying Kai1 gene was amplified, purified and its titer was determined. The rAd-Kai1 was transfected into human laryngeal carcinoma Hep- 2 cells were transfected with empty vector and non-transfected Kai1 laryngeal carcinoma cell lines as control, with methyl thiazolyl tetrazolium (MTT) assay Kai1 on cell proliferation, using RT-PCR technology Transfected cells were tested. Results After rAd-Kai1 was amplified and purified, the titer reached 6 × 1010 PFU / ml. When the viral load was 30 MOI, the transfection rate of Hep-2 cells reached more than 90%. MTT assay showed that the proliferation of Hep-2 cells was not significantly different between transfected empty adenovirus group and untransfected group. The proliferation of Hep-2 cells transfected with Kai1 group was significantly weaker than that of untransfected cells (P <0.05), inhibition rate of up to 29%. Conclusion The tumor metastasis suppressor gene Kai1 / CD82 has an inhibitory effect on the proliferation of laryngeal carcinoma cell lines in vitro.