千金藤素通过调节miR-150和miR-182促进A549细胞凋亡  

Cepharanthine promotes A549 cell apoptosis by regulating miR-150 and miR-182

作　　者：郝瑞敏[1] 谢书阳[1] 韩婉虹 阎云飞[1] 孙允霄[1] 岳真[1] 马颖[1] 李有杰[1] 

机构地区：[1]滨州医学院山东省高校肿瘤分子生物学重点实验室生物化学与分子生物学教研室,山东烟台264003

出　　处：《中国病理生理杂志》2017年第11期1987-1992,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31371321);山东省自然科学基金资助项目(No.ZR2014HQ079;No.ZR2013HL003;No.ZR2014HL056);山东省高校科技计划(No.J13LE11)

摘　　要：目的:探讨千金藤素对人肺癌A549细胞生长的影响及可能机制。方法:千金藤素处理A549细胞后,采用MTT法检测细胞生长抑制率;倒置显微镜观察细胞形态变化;流式细胞术检测细胞凋亡;real-time PCR检测微小RNA(miR)-150、miR-182、p53 mRNA和FOXO1 mRNA的表达变化;通过软件预测miR-150和miR-182的下游靶基因;采用Western blot法分析细胞中p53和FOXO1蛋白表达的变化。结果:在千金藤素作用下,A549细胞生长受到抑制,凋亡率明显增加;千金藤素作用后,细胞内miR-150和miR-182的表达水平显著下降;在10μmol/L的千金藤素作用后,细胞内p53和FOXO1的表达水平升高,而在30μmol/L时,细胞内p53和FOXO1的表达水平降低;在细胞内转染miR-150后,p53表达明显减少,而转染miR-182后,FOXO1表达显著降低。结论:千金藤素能够抑制A549细胞生长,促进A549细胞凋亡,可能是通过抑制miR-150和miR-182表达发挥作用;而miR-150和miR-182可能分别通过下调p53和FOXO1的表达发挥作用。AIM：To investigate the effect of cepharanthine on the growth of human lung carcinoma A549 cells and the underlying mechanism.METHODS：After A549 cells were treated with cepharanthine,the growth inhibitory rate was detected by MTT assay.The cell morphological changes were observed under light microscope.The apoptosis of the A549 cells was analyzed by flow cytometry.The expression of microRNA（miR）-150,miR-182,p53 mRNA and FOXO1 mRNA were detected by real-time PCR.The downstream target genes were predicted by software,and the expression of p53 and FOXO1 was determined by Western blot.RESULTS：After cepharanthine treatment,the growth of A549 cells was inhibited,the apoptosis rate was significantly increased,and the expression levels of miR-150 and miR-182 were significantly decreased.With cepharanthine treatment at 10 μmol/L,the expression levels of p53 and FOXO1 were elevated;however,with cepharanthine at 30 μmol/L,the expression levels of p53 and FOXO1 were decreased.After transfection with miR-150,the expression of p53 was significantly decreased,while the expression of FOXO1 was significantly decreased after transfection with miR-182.CONCLUSION：Cepharanthine inhibits the growth of A549 cells and promotes the apoptosis of A549 cells by inhibiting the expression of miR-150 and miR-182.miR-150 and miR-182 may down-regulate the expression of p53 and FOXO1,respectively.

关 键 词：千金藤素 微小RNA-150 微小RNA-182 P53 FOXO1 

分 类 号：R363[医药卫生—病理学] R285.5[医药卫生—基础医学]