血管紧张素Ⅱ1型受体短发夹环RNA抑制其在哺乳动物细胞中的表达  

Inhibition of angiotensin Ⅱ type 1 receptor expression by short hairpin RNA in mammalian cells

作　　者：孙成林[1] 段志泉[1] 冯宗承[1] 辛世杰[1] 古峻[1] 张京红[1] 张强[1] 

机构地区：[1]中国医科大学附属第一医院血管外科,沈阳110001

出　　处：《中华实验外科杂志》2004年第7期812-814,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30000163)

摘　　要：目的探讨血管紧张素Ⅱ1型受体(AT1R)的短发夹环RNA质粒(pAT1RshRNA)能否抑制AT1R在哺乳动物细胞中的表达。方法构建质粒并转染入鼠血管平滑肌细胞(VSMC)中,24h后应用RTPCR及Westernblot检测VSMC的AT1R的mRNA和蛋白表达变化。结果构建的质粒经测序鉴定验证与预期相符,转染细胞后RTPCR吸光度值比值结果两质粒转染组分别为1.371±0.148和1.453±0.123,与对照组2.088±0.258比较差异有非常显著性(P<0.01);Westernblot结果两质粒转染组分别为1.121±0.037和1.202±0.068,与对照组3.168±0.210比较差异有非常显著性(P<0.01)。提示两质粒均能明显降低AT1R的mRNA和蛋白表达。结论构建的pAT1RshRNA具有RNA干扰作用,能在哺乳动物细胞中抑制该特异基因的表达。Objective To investigate whether the plasmid containing the short hairpin RNA (shRNA) of angiotensin Ⅱ type 1 receptor (AT1R) can inhibit the expression of AT1R in mammalian cells.Methods The plasmids containing the shRNA of AT1R were constructed,and the change of mRNA and protein were detected by RT PCR and Western blot after transfecting vascular smooth musle cells.Results The plasmids was certified to be in the right rank.After transfecting cells,there was significant difference ( P <0.01) in the expression of AT1R mRNA between the gene transfected group (pAT1R shRNA1 1.371±0.148;pAT1R shRNA2 1.453±0.123) and the control group (2.088± 0.258), and there was significant difference ( P <0.01) in the expression of AT1R protein between the gene transfected group (pAT1R shRNA1 1.121±0.037;pAT1R shRNA2 1.202±0.068) and the control group (3.168±0.210).It was indicated that pAT1R shRNA could decrease the expression of AT1R mRNA and protein.Conclusion The plasmids containing the shRNA of AT1R have the RNA interference activity and can inhibit the expression of the specific gene in mammalian cells.

关 键 词：血管紧张素Ⅱ1型受体 短发夹环 RNA 哺乳动物细胞 表达 平滑肌细胞 

分 类 号：Q786[生物学—分子生物学]