靶向Survivin siRNA对乳腺癌MCF-7细胞雌激素受体表达的影响  

Effect of siRNA-targeted Survivin on the Expression of Estrogen Receptor of Breast Cancer MCF-7 Cells

作　　者：黄菁[1] 虞玲[1] 隋俊[1] 张永臣 

机构地区：[1]南京市第二医院检验科,南京210003

出　　处：《华中科技大学学报（医学版）》2016年第5期496-499,503,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81101999)

摘　　要：目的探讨靶向Survivin siRNA对乳腺癌MCF-7细胞雌激素受体表达的影响。方法构建靶向Survivin siRNA载体,按照质粒类型分为空白对照组、阴性对照组和阳性实验组,空白对照组仅加入Lipofectamine 2000,阴性对照组加入空载质粒,阳性实验组加入靶向Survivin siRNA质粒,采用RT-PCR检测Survivin、ER mRNA表达水平,采用Western blot检测细胞Survivin、ER蛋白表达水平,并采用MTT法检测各组对MCF-7细胞增殖抑制率。结果以β-actin作为内参照,空白对照组Survivin、ER mRNA表达量分别为(1.17±0.14)和(0.79±0.11),阴性对照组分别为(1.15±0.12)和(0.80±0.09),阳性实验组分别为(0.43±0.03)和(1.94±0.41),阳性实验组Survivin mRNA表达显著低于空白对照组、阴性对照组,ER mRNA表达显著高于空白对照组、阴性对照组(均P<0.05);空白对照组、阴性对照组Survivin、ER mRNA表达量比较差异无统计学意义(均P>0.05)。Western blot结果显示,阳性实验组Survivin蛋白表达显著低于空白对照组和阴性对照组,ER蛋白表达显著高于空白对照组和阴性对照组。细胞增殖实验结果显示,阳性实验组接种12、24、48h后MCF-7细胞增殖的抑制率逐渐升高,并显著高于同时段空白对照组和阴性对照组(均P<0.05)。结论通过siRNA技术干扰Survivin表达能够显著提高MCF-7细胞雌激素受体表达水平,靶向Survivin siRNA在ER阳性乳腺癌的基因治疗中具有潜在的临床价值。Objective To explore the effect of siRNA-targeted survivin on the estrogen receptor（ER）of breast cancer MCF-7 cells.Methods The siRNA vectors that could target survivin were constructed.Three groups were set up according to different plasmids added：blank control group,in which lipofectamine 2000 was added;negative control group,in which the empty plasmids were given;positive experimental group,in which survivin siRNA was transfected into the cells.mRNA expression levels of survivin and ER were detected by RT-PCR and their protein levels by Western blotting.The inhibition rate of MCF-7 cells proliferation was detected by MTT assay.Results Withβ-actin as an internal reference,the mRNA expression levels of survivin and ER were（1.17±0.14）and（0.79±0.11）in blank control group,（1.15±0.12）and（0.80±0.09）in negative control group,and（0.43±0.03）and（1.94±0.41）in positive experimental group.The mRNA expression levels of survivin were significantly decreased and those of ER increased in positive experimental group as compared with blank control and negative control groups（P〈0.05）.There was no significance in the mRNA expression levels of survivin and ER between the blank control group and the negative control group（P〉0.05）.Similar findings were also observed in the expression of survivin and ER at the protein level.The proliferation inhibition rate was gradually increased from 12,24 to 48 h in the positive experimental group,which was significantly higher than that in blank control and negative control groups at the same time point（P〈0.05）.Conclusion Interfering survivin expression by siRNA technology can significantly enhance the ER levels in MCF-7 cells,suggesting survivin siRNA has potential clinical value in the gene therapy of ER-positive breast cancer.

关 键 词：雌激素受体 小干扰RNA 乳腺癌 细胞增殖 SURVIVIN 

分 类 号：R737.9[医药卫生—肿瘤]