摘要
目的:探讨miR-34a通过下调SIRT1抑制乳腺癌干细胞的机制。方法:使用化学合成的方法合成出miR-34a的mimics或者inhibitors,将其感染乳腺癌MCF-7细胞,通过qRT-PCR的方法验证miR-34a的基因表达。以慢病毒作为载体过表达miR-34a的基因,将能够稳定感染的细胞株筛选出来。设计SIRI1基因中的shRNA,用于干扰乳腺癌MCF-7细胞中的SIRT1的表达。使用qRT-PCR方法和Western blotting技术检测SIRT1中mRNA和蛋白的表达。结果:通过miR-34a的mimics转染的细胞的miR-34a的基因表达水平大于没有转染的细胞,通过miR-34a的inhibitors转染的细胞的miR-34a的基因的表达水平小于没有转染的细胞,差异具有统计学意义(P<0.05)。LV-miR-34a感染细胞可以上调miR-34a的表达,与没有转染的细胞相比,差异具有统计学意义(P<0.05)。shRNA-SIRT1-3组和shRNA-SIRT1-4组中的SIRT1中的mRNA和蛋白的表达出现下降的情况。过表达的miR-34a对SIRT1中的mRNA的表达没有影响。miR-34a的mimics具有抑制SIRT1的蛋白表达的作用,miR-34a的inhibitors具有上调SIRT1的蛋白表达的作用,与没有转染的乳腺癌的MCF-7的细胞相比,差别具有统计学意义(P<0.05)。结论:使用miR-34a的模拟物以及抑制物可以调节miR-34a的表达,从而改变SIRT1的蛋白质的表达。
Objective：To study the mechanism of miR-34a inhibiting breast cancer stem cells by down regulating SIRT1.Methods：The mimics or inhibitors of miR-34a were synthesized by chemical synthesis method,and the MCF-7 cells were infected with qRT-PCR,and the expression of miR-34a gene was confirmed by the method of.Lentiviral vector was used as a vector to express the miR-34a gene,and the cell lines with stable infection were screened out.Design of SIRI1 gene shRNA,used to interfere with the expression of SIRT1 in breast cancer MCF-7 cells.Detection of mRNA and protein expression in SIRT1 by qRT-PCR and Western blotting.Results：No more than the expression level of mimics cells transfected by miR-34a transfected cells miR-34a gene transfected cells,no less than the expression level of inhibitors cells transfected with miR-34a miR-34a gene,the difference was statistically significant（P〈0.05）.LV-miR-34a infected cells can up regulate the expression of miR-34a ,compared with non transfected cells,the difference was statistically significant（P〈0.05）.The expression of mRNA and protein in SIRT1 and shRNA-SIRT1-4 group decreased in the shRNA-SIRT1-3 group.Overexpression of miR-34a had no effect on the expression of mRNA in SIRT1.miR-34a mimics can inhibit the expression of SIRT1 protein,the expression of miR-34a inhibitors is upregulated SIRT1 protein,compared with non transfected breast cancer cells of MCF-7,the difference was statistically significant（P〈0.05）.Conclusion：The expression of miR-34a can be regulated by the miR-34a mimics and inhibitors,which can change the expression of SIRT1 protein.
作者
李晓锋
汪园园
刘希
Li Xiaofeng;Wang Yuanyuan;Liu Xi.(Department of Pathology First Affiliated Hospital of Xi＇an Jiaotong University（ Xi＇an 710061)
出处
《陕西医学杂志》
CAS
2018年第6期683-685,共3页
Shaanxi Medical Journal
基金
陕西省国际科技合作与交流计划项目(2016KW-009)
