RhoA小干扰RNA对胃癌AGS细胞基因表达谱的影响

目的：应用RhoA小干扰RNA（small interfering RNA，siRNA）转染胃癌细胞系AGS细胞，采用高通量基因芯片技术，筛选基因表达谱的变化，为进一步研究RhoA信号转导通路奠定基础。方法：用pSilencer3．1载体构建RhoA／siRNA重组质粒。Effectene转染试剂转染AGS细胞，空载体pSilencer3．1转染对照组。应用G418筛选转染细胞系，获得稳定转染细胞；美国Agilent基因芯片检测细胞基因表达谱变化。结果：基因芯片检测全基因组，发现表达上调的有1151个基因，表达下调的有1079个基因，涉及信号转导、细胞周期、细胞凋亡等相关信号分子。生物信息学分析显示，与细胞凋亡有关的caspase家族，以及细胞周期调控基因存在明显的差异表达。结论：基因芯片结果提示干扰RhoA表达后，与凋亡和增殖有关的分子在RhoA介导的肿瘤细胞异常增殖信号转导中起着重要的调控作用，为下一步研究其信号调控网络机制提供重要的信息。

The alteration of gene expression profile in gastric cancer AGS cells transfected with RhoA small interfering RNA

Objective ：To analyse the alterations of gene expression in gastric cancer cell line AGS transfected with RhoA small interfering RNA （siRNA） by DNA gene microarray technology. Methods： The recombinant RhoA/siRNA plasmid pSilencer 3.1 was constructed and transfected into gastric cancer cell line AGS with effectene reagent. pSilencer 3.1 was transfected with the same cell line as the control . The two groups were subjected to G418 selection. The gene expression profile was analyzed by Agilent Gene Chip. Results ： The gene microarray chip demonstrated that 1151 genes were up -regulated and 1079 genes down -regulated, which involved in signal transduction, cell cycle , apoptosis et al. Bioinformation analysis showed that the caspase family which related to cell apoptosis, the cell cycle related molecules,were important in RhoA mediated signal transduction. Conclusion： The gene microarray is a good method with advantages of high flux and high efficiency , which brought some new clues to study the molecular mechanism of RhoA signaling in cancer .