c—myc靶向小干扰RNA诱导乳腺癌细胞
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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
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