c—myc靶向小干扰RNA诱导乳腺癌细胞

翟荣林王国斌 夏泽峰 【摘要】 耳的构建小干扰RNA(siRNA)表达载体，体外观察其诱导乳腺癌细胞凋亡效应。 方法基因克隆技术构建人c—mye癌基因特异性siRNA表达载体，体外转染MCF．7。48 h后检测c—my茈基因表达状况并观察MCF一7凋亡诱导效应。结果 (1)成功构建siRNA表体载体：pEGFP—C1／U6．1、pEGFP—c1／U6—2和pEGFP—C1／U6．3；(2)siRNA表达载体转染MCF一7 48 h后。pEGFP-C1／U6．2组显著抑制胞内c．myc表达(80％)；(3)转染24 h和48 h后，pEGFP．C1／U6—2组凋亡率分别为11．01％和21．30％。明显高于对照组(P<0．01)。结论构建的c—myc靶向siRNA表达载体能显著下调c．myc在MCF-7中的过表达。诱导乳腺癌细胞株MCF-7凋亡。 【关键词】 c．myc基因； siRNA； 乳腺癌； 脱噬作用 Apoptasis induction in breast~ancef cells by siRNA targeting gene c-myc 【Abstract】 Objective To observe apoptoais induction in breast cancer cells by siRNA．Methods The siRNA expression vectors were constructed and transfected into MCF一7．At 48 h after transfection． cell apoptosis in MCF-7 was detected by FCM ．The gene expression of C-ITIyc was detected bv RT．PCR and Western blotting respectivdy．Results (1)Transfectbn of pEGFP—C1／U6—2 group into MCF-7 down-regulated C-myc expression level greatly at 48 h after transfection(80％)；(2)The apoptosis per—centage of pEGFP—C1／U6．2 group was 11．01％ at 24 h and 21．30％ respectivdy at 48 h after transfec． tion。significantly higher than in the control group(P<0．01)．Conclusion The siRNA expression vec—tor we comt~ct can effectively down-regulate c-myc over expression and remarkably induce apoptosis in MCF一7 cell line． 【Key words】c-myc gene； siRNA； Breast carcinoma； Apoptosis