RNA干扰介导的结肠癌细胞中血管内皮生长因子基因沉默
RNA interference-mediated gene silencing of vascular endothelial growth factor in colon cancer cells.
作者信息
Li Tie-Jun, Song Jian-Ning, Kang Kai, Tong Shu-Sheng, Hu Zan-Lan, He Tong-Chuan, Zhang Bing-Qiang, Zhang Cai-Quan
机构信息
Department in General Surgery, the First Affiliated Hospital, Chongqing University of Medical Science, Chongqing 400016, China.
出版信息
World J Gastroenterol. 2007 Oct 28;13(40):5312-6. doi: 10.3748/wjg.v13.i40.5312.
AIM
To inhibit the expression of vascular endothelial growth factor (VEGF) in colon cancer cell line by RNA interference (RNAi).
METHODS
Followed the service of E-RNAi, we designed and constructed two kinds of shRNA expression vectors aiming at the VEGF gene, then transfected them into colon cancer HT29 cells by lipofectamine(TM) 2000. The level of VEGF mRNA was investigated by RT-PCR and Northern blotting. The protein expression of VEGF was observed by immunofluoresence staining and Western blotting.
RESULTS
We got two kinds of VEGF specific shRNA expression vectors which could efficiently inhibit the expression of VEGF in HT29 cells. RT-PCR, Northern blotting, immunofluoresence staining and Western blotting showed that inhibition rate for VEGF expression was up to 42%, 89%, 73% and 82%, respectively.
CONCLUSION
The expression of VEGF can be inhibited by RNA interference in HT29 cells.
目的
通过RNA干扰（RNAi）抑制结肠癌细胞系中血管内皮生长因子（VEGF）的表达。
方法
按照E-RNAi服务，设计并构建针对VEGF基因的两种shRNA表达载体，然后用脂质体2000将它们转染到结肠癌HT29细胞中。通过RT-PCR和Northern印迹法研究VEGF mRNA的水平。通过免疫荧光染色和Western印迹法观察VEGF的蛋白表达。
结果
我们获得了两种VEGF特异性shRNA表达载体，它们可以有效抑制HT29细胞中VEGF的表达。RT-PCR、Northern印迹法、免疫荧光染色和Western印迹法显示，对VEGF表达的抑制率分别高达42%、89%、73%和82%。
结论
RNA干扰可在HT29细胞中抑制VEGF的表达。
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