摘要
目的研究CTGF在人食管癌细胞中的表达情况及其对人食管癌细胞系Eca109细胞凋亡的影响和可能机制。方法应用Real Time PCR获取编码CTGF全长的cDNA,经免疫组织化学染色,经Western Blot检测食管癌中CTGF mRNA及蛋白的表达情况;应用pcDNA3.1质粒和慢病毒载体分别上调和下调Eca109细胞中CTGF的表达,应用流式细胞仪检测细胞凋亡比率;Western Blot检测凋亡相关蛋白BCl-XL表达。结果与正常组织比较,食管癌组织中CTGF mRNA及蛋白的表达水平均明显增高,免疫组化结果显示食管癌标本中CTGF显色较正常对照组织强。转染pcDNA3.1 CTGF质粒后,有效增强了Eca109细胞中CTGF蛋白的表达和凋亡的抵抗作用,且抗凋亡蛋白BCl-XL表达水平上调。用RNAi逆转录病毒转染后,有效下调了内源CTGF的表达,对INF-γ介导的凋亡抵抗作用较对照组下降,与对照组相比,BCl-XL蛋白表达水平亦下调。结论在食管癌中CTGF表达水平明显增高,并且可以通过上调抗凋亡蛋白BClXL表达从而抑制细胞凋亡。
Objective To observe the CTGF expression in the sample of ESCC and study the effects and mechanism of CTGF on cell apoptosis of Eca109 cell. Methods Application of Real Time PCR to obtain full-length cDNA encoding CTGF,after immunohistochemical staining,the expression of CTGF Western Blot for the detection of esophageal cancer specimens in mRNA and protein; the application of pcDNA3. 1 plasmid and lentiviral vectors were up-regulated and down regulated CTGF expression in Eca109 cells,flow cytometry was used to detect the expression of apoptosis ratio; Western Blot for the detection of apoptosis related protein BCl-XL. Results Compared with the control,the expression level of CTGF mRNA and protein in ESCC were significant higher. In cancerous tissues,CTGF immunoreactivity was more intense than in corresponding normal tissues. The expression of CTGF protein and the expression of BCl-XLwere increased in Eca109 cells after transfeceting with CTGF expression vector. Western blot results showed that siRNA system markedly decreased the basic expression level of CTGF compared with the control sequence. The knock down of CTGF promotes cell apoptosis and the expression of BCl-XLwas lower compared with the control. Conclusion Overexpression of CTGF occurs in a significant proportion of esophageal squamous cell carcinoma samples and enhance the resistance of apoptosis through up-regulation of BCl-XL.
出处
《实用癌症杂志》
2014年第6期623-625,共3页
The Practical Journal of Cancer
