摘要
瞄准：由 RNA 干扰(RNAi ) 在结肠癌房间线禁止脉管的内皮生长因素(VEGF ) 的表示。方法：跟随了 E-RNAi 的服务，我们设计了并且构造了二种 shRNA 表示向量瞄准 VEGF 基因，然后 transfected 他们进由 lipofectamineTM 的结肠癌 HT29 房间 2000。VEGF mRNA 的水平被 RT-PCR 并且北弄污调查。VEGF 的蛋白质表示被 immunofluoresence 染色并且西方的弄污观察。结果：我们得到了能高效地在 HT29 房间禁止 VEGF 的表示的二种 VEGF 特定的 shRNA 表示向量。RT-PCR，北弄污， immunofluoresence 染色并且西方的弄污证明为 VEGF 表示的那抑制率分别地直到 42% ， 89% ， 73% 和 82% 。结论：VEGF 的表示能被 RNA 干扰在 HT29 房间禁止。
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 lipofectamineTM 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.
基金
Supported in part by research grants from the National Natural Science Foundation of China,No.30300298
the National Natural Science Foundation of China's Joint Research Fund for Overseas Chinese Young Scholars,No.30228026
关键词
RNA干涉
血管
结肠癌
细胞生长
RNA interference
Vascular endothelialgrowth factor
Colon cancer
Northern blotting
Western blotting
