Combined use of RT-PCR and gel electrophoresis to detect expression of transforming growth factor beta1 in mouse lung fibroblasts in vitro

VernacularTitle:RT-PCR联合凝胶电泳成像技术检测体外培养小鼠肺成纤维细胞转化生长因子β1的表达
Author: Weitao YU ; Dongjian WANG ; Lingyan AI
From: Chinese Journal of Tissue Engineering Research 2017;21(8):1268-1272
CountryChina
Language:Chinese
Abstract: BACKGROUND: As a combination of reverse transcription (RT) and polymerase chain reaction (PCR), RT-PCR has been used to detect gene expression levels in cells and tissues, RNA virus contents in cells and specific gene cloned cDNA sequences.OBJECTIVE: To detect the inhibitory effcet of Stealth siRNAs on the expression of transforming growth factor β1 (TGF-β1).METHODS: There were blank control, empty vector transfection, stealth-48, stealth-166, and stealth-594 groups. Three stealth siRNAs aimed at different sequences in TGF-β1 mRNA were made, and were then transfected into BALB/c mouse lung fibroblasts in vitro. The expressions of TGF-β1 and connective tissue growth factor were detected by RT-PCR.RESULTS AND CONCLUSION: In different time periods, the TGF-β1 expression was differentially depressed by three stealth siRNAs, especially stealth-166. The inhibitory effects varied with time, which could be detective at 48 hours,reached the peak at 72 hours and then began to attenuate at 96 hours. Our findings show that the inhibitory effect of stealth siRNAs on the TGF-β1 expression in mouse lung fibroblasts can be detected by RT-PCR.