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

发布时间：2018-01-23 08:23

本文关键词： RNA 小分子干扰转化生长因子β 成纤维细胞组织工程组织构建 Stealth siRNA 肺成纤维细胞抑制　出处：《中国组织工程研究》2017年08期 　论文类型：期刊论文

【摘要】：背景:RT-PCR技术是将RNA的反转录和cD NA的聚合酶链反应相结合的技术。常用于检测细胞组织中基因表达水平、细胞中RNA病毒的含量和直接克隆特定基因的cD NA序列等。目的:运用RT-PCR技术检测Stealth siRNA对转化生长因子β1的抑制作用。方法:实验分为空白对照组、空转染组、stealth_48组、stealth_166组和stealth_594组。针对BALB/c小鼠转化生长因子β1基因组,选择不同位点设计3套siRNA基因序列,转染体外培养的小鼠肺成纤维细胞,RT-PCR法检测其对转化生长因子β1和下游结缔组织生长因子表达的影响。结果与结论:(1)RT-PCR结果显示3种Stealth siRNA对转化生长因子β1表达在不同时间有不同程度的抑制作用,以Stealth_166效果更为明显;抑制效果与转染时间长短相关,48 h后就可检测出明显抑制,72 h达到最高峰,96 h后开始减弱;(2)结果说明,RT-PCR技术可用于特异性Stealth siRNA抑制转化生长因子β1在小鼠肺成纤维细胞的表达效果的检测。
[Abstract]:Background: RT-PCR is a combination of reverse transcription of RNA and polymerase chain reaction of cdna. It is often used to detect gene expression level in cell tissues. The content of RNA virus in cells and the cdna sequence of specific genes were cloned directly. Objective: to detect Stealth by RT-PCR technique. The inhibitory effect of siRNA on transforming growth factor 尾 1. Methods: the experiment was divided into blank control group. The genome of transforming growth factor 尾 1 (TGF- 尾 1) in BALB/c mice was detected in the empty transfection group and the stealth166 group and stealth_594 group. Three sets of siRNA gene sequences were designed at different sites and transfected into mouse lung fibroblasts in vitro. The effect of TGF- 尾 1 and downstream connective tissue growth factor (TGF- 尾 1) on the expression of TGF- 尾 1 and connective tissue growth factor (TGF- 尾 1) was detected by RT-PCR. RT-PCR results showed that the expression of transforming growth factor 尾 1 was inhibited by three kinds of Stealth siRNA at different times. The effect of Stealth_166 was more obvious. The inhibitory effect was correlated with the length of transfection time. After 48 h, it was found that the inhibitory effect was obviously inhibited at 72 h and reached the peak at 96 h, then it began to weaken. The results indicated that RT-PCR could be used to detect the expression of transforming growth factor 尾 1 in mouse lung fibroblasts by specific Stealth siRNA.
【作者单位】：解放军第184医院;
【基金】：南京军区医药卫生科研基金资助项目(07M057)~~
【分类号】：R-33
【正文快照】： 引用本文:余维涛,汪东剑,艾凌艳.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[J].中国组织工程研究,2017,21(8):1268-1272. 文章快速阅读:Stealth si RNA对小鼠肺成纤维细胞转化生长因子β1 m RNA表达的影响评价:Stealth引物设计si RNA对

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