摘要
目的探讨去甲基化药物5-氮杂-2’-脱氧胞苷(简称5-Aza)对转录调节因子Snail的作用,转染反义Snail与5-Aza对上皮间质转化相关膜蛋白E-钙粘素的mRNA水平的影响及二者联合作用的可能机制。方法应用MTT、PCR、实时荧光定量PCR、检测低分化胃腺癌细胞株分别转染反义Snail、应用5-Aza单独或协同干预后对细胞的生长增殖及Snail、E-钙粘素mRNA表达的影响。结果与空白对照组比较5-Aza干预、转染反义Snail分别及二者共同干预均增加E-cadherin的mRNA的表达,空质粒组无显著差异。5-Aza上调Snail基因mRNA的表达。两种干预方式处理均能抑制胃癌细胞的生长。结论 5-Aza、转染反义Snail均使E-cadherin mRNA表达增强,且药物作用呈时间浓度依赖性。5-Aza能上调Snail基因的表达,提示甲基化作用在同时下调转录调节子Snail基因表达和抑制E-钙粘素表达。
Objective The aim of the research is to study the effect of demethylation drug 5- Aza on E-cadherin transcription factor Snail, and discuss the possible mechanism of joint effect of antisense Snail and 5-Aza on up-regulate the epithelial adhesion factor Ecadherin mRNA levels and the influence of the two. Methods Applying MTr, PCR and real-time fluorescent quantitative PCR to detect poorly differentiated gastric adenocarcinoma cell lines transfection of antisense Snail, and observing, the effects of using drug 5-Aza alone or combined with antisense Snail synergistically on gastric cancer cell proliferation and Snail and E-cadherin mRNA expression. Results Compared with the blank control group, 5-Aza intervention, transfection of antisense Snail respectively and two joint intervention all increased the expression of E-cadherin mRNA, and there was no significant difference in empty plasmid group. 5-Aza upregulated the expression of Snail gene mRNA. Both kinds of intervention treatment can inhibit the growth of gastric carcinoma cells. Conclusion Both 5-Aza and transfection of antisense Snail can increase the expression of E-cadherin mRNA, and the drug effect is dependent on time and concentration. 5-Aza can up-regulate the expression of Snail gene and suggest methylation, at the same time, it down-regulates transcriptional regulator Snail gene expression and inhibits the expression of E- cadherin.
出处
《中外医疗》
2013年第22期4-5,共2页
China & Foreign Medical Treatment
