利用RNA干扰技术降低REV3基因在人类结肠癌细胞(SW480)中的表达,以荧光实时定量PCR检测REV3表达量的降低情况,选择低表达效率具有统计学意义的细胞作为实验组细胞。运用细胞生长曲线、MTT、微核和姐妹染色单体交换等方法,对实验组和对照组细胞进行细胞生长周期、增殖变化情况和遗传信息表达等指标的检测。结果显示:REV3低表达的结肠癌实验组细胞在细胞增殖以及细胞的微核和姐妹染色单体交换等遗传信息表达均明显低于结肠癌对照组细胞,实验结果具有统计学意义(P〈0.05);结肠癌的两对照组间(阴性和空白)的结果虽然有一定的差异,但没有统计学意义。研究结果提示,REV3低表达时,可能对结肠癌细胞(SW480)的生长与增殖产生影响,并对微核和姐妹染色单体交换等遗传不稳定现象的产生有一定的抑制作用。
To investigate the roles of human REV3 gene in proliferation and genomic stability,we experimentally suppressed the REV3 expression in human colon cancer cells(SW480) by the interference RNA technology(RNAi) as moni-tored by real-time RT-PCR.Compared to untreated or mock-treated cells,ablation of REV3 significantly reduced cell growth rate,micronucleus formation and the frequency of sister chromatid exchange.Whereas the differences between untreated and mock-treated controls were insignificant.Indicators of cell cycle,proliferation and the expression of genetic information in cells of case group,which displayed lower-level expression of REV3,were extremely lower than the control group,and the difference was significant(P0.05).Differences in the two control groups were not significant.This suggested that the reduced expression of REV3 may affect the growth and proliferation of colon cancer cells(SW480),and,to some extent,contribute to suppression of the genetic instability occurred in micronuclei and sister chromatids.Based on the results from this study,REV3 plays an important role in different cellular growth periods and physiological conditions,and its differential expression directly affects the development of human colon cancer cells.