Combined use of RT-PCR and gel electrophoresis to detect expression of transforming growth factor beta1 in mouse lung fibroblasts in vitro / 中国组织工程研究

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.

Inhibitive effect of specific stealth siRNAs on TGF-?_1 expression of mouse lung fibroblasts / 第三军医大学学报

Objective To design and produce specific stealth siRNAs and detect their inhibitive effects on TGF-?1 expression.Methods Three stealth siRNAs aimed directly at different sequences(stealth-48,stealth-166,stealth-594) in TGF-?1 mRNA were made.These siRNAs were transfected into BALB/c mouse lung fibroblast in vitro,then the TGF-?1 expression in those lung fibroblasts was detected by immunohistochemistry.Results In different time periods,the TGF-?1 expression was differently depressed by three stealth siRNAs in vitro.The inhibitory effects of stealth-166 was better than the other two stealth siRNAs.The inhibition could be detected in 48 h,reached the highest level in 72 h and began to attenuate 96 h later.Conclusion The TGF-?1 expression of mouse lung fibroblasts in vitro can be depressed by synthetic stealth siRNAs.It seems possible to provide a new way for the treatment of pulmonary interstitial fibrosis.