RNA干扰技术抑制EMMPRIN表达对人绒癌细胞JEG-3侵袭性的实验研究  

Study on the RNAi inhibit expression of EMMPRIN and invasion of JEG-3

作　　者：王永清[1] 赵扬玉[1] 江元慧[1] 杨硕[1] 

机构地区：[1]北京大学第三医院妇产科,100083

出　　处：《中国优生与遗传杂志》2008年第2期28-30,共3页Chinese Journal of Birth Health & Heredity

摘　　要：目的研究RNA干扰技术抑制人绒癌JEG-3细胞EMMPRIN的表达,探讨EMMPRIN在滋养细胞侵袭行为中的作用。方法设计、体外化学合成靶向EMMPRIN的短发夹状双链RNA(shRNA),分别与脂质体LipofectamineTM2000结合后转染体外培养的人绒癌细胞系JEG-3。分为A实验组:每组中加入1μg重组质粒和5μl阳离子脂质体Metafectene;B阴性对照组:只加5μl脂质体转染试剂;C无关对照组(无关dsRNA对照组,载体试剂盒自带)。三组细胞孵育48h后,Western blot和RT-PCR技术检测EMMPRIN蛋白和mRNA的表达,明胶酶谱测定MMP2,9的表达。结果RNA干扰技术可以显著的靶向抑制EMMPRIN基因在人绒癌细胞中的表达,与B组和C组相比,A组细胞EMMPRIN蛋白和EMMPRINmRNA表达分别减少了75.2%和62.3%(P<0.01);B、C组EMMPRIN蛋白和EMMPRIN mRNA表达分别减少了3.3%、2.4%和3.2%、1.8%(P>0.05);A组细胞在转染后的12,24,36,48小时MMP2的分泌分别下降了13.5%,29.6%,58.3%和66.6%;MMP9的分泌分别下降了10.7%,33.8%,47.5%和62.6%,相邻组间比较差异显著(P<0.05)。结论EMMPRIN与滋养细胞的侵袭功能密切相关,抑制滋养细胞中EMMPRIN的表达可以抑制滋养细胞的侵袭。Objective： To investigate the role of RNAi in the inhibition expression of EMMPRIN in JEG - 3 and the role of EMMPRIN in invasion of cytotrophoblast. Methods： The double standed RNA （dsRNA） with hairpin structures targeted specifically at EMMPRIN mRNA, were designed, synthesized. These siRNA were transfected into the cultured JEG -3 after combined with Lipofetamine TM2000. The trial had been divided into 3 group： experiment group A： 5txl of lipoplast and Metafectene/1μg recombinant plasmid were added into each poreof; negative control group B： 5μl of Lipofectamine MT transfection was added into negative control group; and blank control group C （Interfered by no factors）. RT - PCR and Western bolt were used to detect the expression of mRNA in EMMPRIN as well as the protein respectively before transfection and 48 hours after transfection, Gelatin Zymography detect the expression of MMP2, 9. Results ： RNAi can significantly inhibit the expression of EMMPRIN mRNA and protein. The expression of EMMPRIN mRNA and protein of A group was gradually decreased 75. 2% and 62. 3% than group B and C （ P 〈0. 01 ） ; the expression of EMMPRIN mRNA and protein of B and C group was gradually decreased 3.3%, 2.4% and 3.2%, 1.8% （ P 〉0. 05） ; A group at 12, 24, 36 and 48 hours after transfection gradually decreased the secretion of MMP2 13.5% , 29. 6%, 58. 3% and 66.6% and MMP9 decreased 10. 7%, 33.8%, 47. 5% and 62.6%, while there were significant differences between the near double groups （ P 〈0. 05）. Conclusion： EMMPRIN is correlated with invasion of Tropboblast. To inhibit the expression of EMMPRIN in trophoblast can inhibit the invasion of trophoblast.

关 键 词：RNA干扰技术 细胞外基质金属蛋白酶诱导因子 绒癌JEG-3细胞系 侵袭力 

分 类 号：R737.33[医药卫生—肿瘤]