摘要
目的探讨转染p21WAF1/CIP1基因(p21基因),对宫颈癌Hela细胞生长抑制作用及p21基因转染Hela细胞对顺铂敏感性的影响。方法应用脂质体转染技术,将质粒pcDNA3转入人宫颈癌Hela细胞中,经G418筛选,转染阳性克隆细胞连续传代培养,并采用中性红摄入法测定顺铂对p21基因转染人宫颈癌Hela细胞增殖抑制作用,流式细胞仪(FCM)检测细胞周期变化。结果经G418筛选,Hela细胞全部死亡,转染质粒pcDNA3的Hela细胞存活,并可连续传代。p21基因转染后,细胞生长明显受到抑制,细胞周期停滞于G1期,与对照组相比,细胞周期G1期比例明显增加。同时p21表达阳性的Hela细胞对顺铂敏感性增强。结论p21基因转染能使Hela细胞出现生长抑制并对顺铂化疗作用敏感性明显增强。
Objective To investigate the effects of human p21^WAF1/GIP1（p21 gene）transfection on the growth and ehemosensitivity of human cervical carcinoma cell line hela. Methods Recombined vector peDNA3 were transferred to Hela cell line by Lipofection. two een lines were selected by G418,and the selected cell lines were subeultured. Hela een ;line were enclosed as control group and the Neutral red was used to explore the inhibitory effect of chemotherapeutic drugs on human Hela cell transfected with p21 gene and the changes of cell cycle and een apoptosis rate were detected by flow cytometry. Results After G418 selecting,Hela cell line in control group was died,and the cell line transferring peDNA3 were subcultured. After transection of p21 gene,cdl growth was obviously inhibited and resulted in an accumulation of cell in G1 phase, presenting that the proportion of the cell in G1 phase was obviously increased and that of the cells in S and G2/M phases was obviously decreased. Respectively as compared with control group. The apoptotic index was increased in Hela-p21 cell.p21-positive Hela was more sensitive to Cisptatin. Conclusion The exogenous p21 gene expressions not only induced great suppression of cell growth but also increased chemosensitivity of human cervical carcinoma.
出处
《中国实验诊断学》
2006年第8期897-899,共3页
Chinese Journal of Laboratory Diagnosis
