摘要
目的构建siRNA稳定表达载体,抑制TIEG基因在瘢痕疙瘩成纤维细胞中的表达,为研究TIEG在瘢痕疙瘩等纤维化疾病发生中的作用提供有效的实验工具。方法根据siRNA设计原则设计并合成带有双向启动子具有转录功能的siRNA载体,转入成纤维细胞内,干扰TIEG基因的表达,利用载体可以发荧光观察转染效果,反转录聚合酶链反应(RT-PCR)检测siRNA对TIEG基因的抑制效果。结果瘢痕成纤维细胞经RNAi沉默后TIEGmRNA表达下调。结论成功建立siRNA载体,并在瘢痕疙瘩成纤维细胞中抑制TIEG基因的表达。
Objective To silence the expression of TIEG in keloid fibroblast by stable expression of vectored siRNA and to provide an experiment tool for functional research of TIEG in fibrosis. Methods According to the principles of siRNAdesign, vectored siRNA with bidirectional promoter and transcription function was synthesized, and transfected into keloid fibroblast to silence the expression of TIEG. The effect of transfection was observed with fluorescence.The interference on TIEG mRNA expression was detected by RT-PCR. Results The expression of TIEG mRNA in keloid fibroblast was decreased after RNAi. Conclusion SiRNA vector was constructed successfully and found to be active in silencing of TIEG expression in keloid fibroblasts.
出处
《中国药物与临床》
CAS
2007年第2期106-108,共3页
Chinese Remedies & Clinics
基金
山西省青年科技研究基金资助项目(2006021047)
