miR-21-5p靶向调控Spry1影响人乳腺癌细胞ERK的磷酸化  

miR-21-5p modulates ERK phosphorylation by targeting Spry1 in breast cancer cells

作　　者：孙方正 檀军 陆祥 陈伟 Sun Fangzheng;Tan Jun;Lu Xiang;Chen Wei(Department of Histology and Embryology,Zunyi Medical University,Zunyi Guizhou 563099,China)

机构地区：[1]遵义医科大学组织胚胎学教研室,贵州遵义563099

出　　处：《遵义医科大学学报》2021年第1期21-27,共7页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:81460466);贵州省科学技术基金资助项目(NO:黔科合J字[2011]2122)。

摘　　要：目的探讨miR-21-5p在人乳腺癌细胞中调控靶基因影响ERK磷酸化的分子机制。方法利用生物信息学预测miR-21-5p潜在靶基因,双荧光素酶报告实验证实miR-21-5p与预测基因之间的靶向关系;分别转染miR-21-5p mimic和inhibitor及相应阴性对照后,通过RT-qPCR和Western blot检测目的基因的表达。结果Targetscan-human 7.1数据库显示Spry1是miR-21-5p潜在的靶基因;双荧光素酶报告实验显示,与阴性对照组相比,miR-21-5p mimic和psiCHECK2-Spry1-WT共转组荧光素酶强度明显降低(P<0.001)。MCF-7细胞中过表达miR-21-5p,Spry1 mRNA表达无明显变化(P>0.05),Spry1蛋白表达降低(P<0.05),ERK磷酸化水平升高(P<0.05);抑制miR-21-5p表达,Spry1 mRNA表达无明显变化(P>0.05),Spry1蛋白表达升高(P<0.05),ERK磷酸化水平降低(P<0.05)。过表达Spry1后,ERK磷酸化水平降低(P<0.05);干扰Spry1表达,ERK磷酸化水平升高(P<0.05)。结论在人乳腺癌细胞中Spry1是miR-21-5p的靶基因,在转录后水平或翻译水平miR-21-5p可以抑制Spry1的表达,促进ERK的磷酸化。Objective To investigate the molecular mechanism of miR-21-5p regulating target genes and affecting ERK phosphorylation in human breast cancer cells.Methods Bioinformatics was used to predict the potential target gene of miR-21-5p.Luciferase reporting experiment confirmed the targeting relationship between miR-21-5p and the predicted gene.After transfection of miR-21-5p mimic and inhibitor and corresponding controls respectively,the expression of the target gene was detected by RT-qPCR and western blot assay.Results Targetscan-human 7.1 database showed that Spry1 was a potential target gene for miR-21-5p.The luciferase reporter experiment showed that the luciferase intensity in miR-21-5p mimic and psiCHECK2-Spry1-WT conversion group decreased compared with the control group.Overexpression of miR-21-5p did not change the expression of Spry1 mRNA in MCF-7 cells,but the expression of Spry1 protein decreased and the level of ERK phosphorylation increased.When the expression of miR-21-5p was inhibited,the expression of Spry1 mRNA had no significant change and the expression of Spry1 protein increased,and the level of ERK phosphorylation decreased.After overexpression of Spry1,ERK phosphorylation level decreased.When the expression of Spry1 was interfered,the phosphorylation level of ERK was increased.Conclusion Spry1 is the target gene of miR-21-5p in human breast cancer cells.miR-21-5p could inhibit the expression of Spry1 and promote the phosphorylation of ERK at post-transcriptional or translation levels.

关 键 词：乳腺癌 miR-21-5p Spry1 ERK磷酸化 

分 类 号：R737.9[医药卫生—肿瘤]