紫杉醇对人肝癌SMMC7721细胞NDRG1表达及增值的影响

Paclitaxel Effect on NDRG1 Expression and Proliferationin in Hepatic Carcinoma Cell SMMC7721

旨在探索紫杉醇对人肝癌SMMC7721细胞NDRG1表达的影响,及紫杉醇对肝癌SMMC7721细胞系增殖的抑制作用。分别提取紫杉醇处理前后SMMC7721细胞的RNA,进行逆转录-聚合酶链反应(RT-PCR),判断紫杉醇处理前后肝癌细胞中NDRG1表达的情况;采用蛋白印迹技术(Western blotting)分析紫杉醇处理前后肝癌细胞中NDRG1蛋白表达的情况;应用不同浓度紫杉醇处理肝癌细胞,以MTT法检测处理前后肝癌细胞的抑制率,流式细胞术(FCM)观察细胞周期变化的况。结果表明紫杉醇处理后的肝癌SMMC7721细胞中NDRG1表达下降,紫杉醇浓度越高,NDRG1表达水平越低,具有浓度依赖性。以MTT法观察紫杉醇对肝癌细胞的抑制作用,试验结果表明不同浓度的紫杉醇处理肝癌SMMC7721细胞后,癌细胞被明显抑制;以流式细胞术观察紫杉醇作用后肝癌SMMC7721细胞周期的变化,结果显示G2-M期细胞比例升高的程度随浓度增高而升高,细胞越来越多地被阻滞在G2-M期,不能继续分裂增殖。分化相关基因NDRG1的表达可能是肝癌的发病机制之一,紫杉醇可抑制肝癌SMMC 7721细胞中NDRG1的表达;同时紫杉醇能使肝癌SMMC7721细胞的生长阻滞在G2-M期,从而显著抑制SMMC7721细胞的增殖,并且具有剂量、时间依赖效应。

It was to investigate the paclitaxel effect on differentiation related gene NDRG1 expression in hepatic carcinoma SMMC7721 cells,and the depressant effect in hepatic carcinoma cell SMMC7721 proliferation.Hepatic carcinoma SMMC7721 cell RNA was extracted before and after dealt with paclitaxel,and the NDRG 1 expression in hepatic carcinoma cells was detected by RT-PCR.Western blotting assay was used to determine expression levels of NDRG1;SMMC7721 cells were treated with paclitaxel,cell proliferation was detected by MTT assay,and cell cycle was detected by fIow cytometry（FCM）.The NDRG1 expression decreased in hepatic carcinoma cell SMMC 7721 after dealt with paclitaxel through RT-PCR and Western blotting.The experiments results indicated that the hepatic carcinoma cells were significantly inhibited after the different paclit axel densities by MTT.The result by FCM showed that paclitaxel can lengthen the G2-M phase in SMMC7721 cells.The NDRG1 gene expression could be one of the hepatic carcinoma pathogenesis,and paclitaxel could inhibited the NDRG1 expre ssion in hepatic carcinoma SMMC7721 cells.At the same time,paclitaxel could inhibit the hepatic carcinoma SMMC7721 cells growth in G2-M term,thus,inhibit the cell multiplication significantly,and displays the relying effect of doses and time.