miRNA-350靶向MAPK14及JNK调控H9c2细胞增殖和凋亡  

Regulation of miRNA-350 on proliferation and apoptosis of H9c2 cell through targeting MAPK14 and JNK gene

作　　者：张淑华[1] 洪浪[1] 王云霞[1] 吴志婷[1] 刘秋玲[1] 葛郁芝[1] ZHANG Shu-Hua;HONG Lang;WANG Yun.Xia(Jiangxi Provincial People＇s Hospital,Jiangxi Provincial Institute of Cardiovascular Disease,Nanchang 330006,Jiangxi,China)

机构地区：[1]江西省人民医院江西省心血管病研究所,江西南昌330006

出　　处：《中国老年学杂志》2018年第18期4482-4485,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(81260031);江西省卫生计生委科技计划项目(20181014)

摘　　要：目的观察miRNA(miR)-350对H9c2细胞凋亡的诱导作用。方法 H9c2细胞转染miR-350诱导细胞肥大。显微镜下观察细胞形态变化并测量细胞表面积;运用MTT法分析细胞增殖,流式细胞技术检测细胞凋亡。并采用Western印迹法检测靶基因丝裂原活化蛋白激酶(MAPK) 14及c-Jun氨基末端激酶(JNK)的蛋白表达,逆转录聚合酶链反应(RT-PCR)的方法检测MAPK14及JNK m RNA表达。结果 miR-350转染第7天细胞发生明显肥大和皱缩,miR-350及sh-MAPK14均能诱导细胞肥大;MTT结果显示miR-350组细胞活性显著下降(P<0. 05); sh RNA-JNK/p38组细胞活性抑制最为显著(P<0. 01);流式细胞术检测结果表明:与对照组比较,转染sh RNA-JNK/p38组和转染miR-350组细胞凋亡率升高,分别是16. 36%和8. 39%。miR-350可在转录水平抑制内源性p38和JNK基因的表达。然而,miR-350上述活性均可被miR-350 antagomir有效阻滞。结论 miR-350靶向MAPK14及JNK基因调控H9c2细胞的增殖和凋亡。Objective To investigate the effect of miR-350 on apoptosis of H9 c2 cell.Methods H9 c2 cells were transfected with miR-350 over-expression to induce cell hypertrophy. Then cells were observed in cell morphology and the area of the individual H9 c2 cell was measured. The proliferation of cell was analyzed by MTT,and apoptosis was detected by flow cytometry. The m RNA and protein expressions of target genes MAPK14 and JNK were detected by Western blot and RT-PCR,respectively.Results Seven days after transfection with miR-350,the area of the individual H9 c2 cell was significantly increased,the activity of cells transfected with miR-350 was decreased（ P〈0.05）and the inhibition of cell activity was most significant in sh RNA-JNK/P38 group（ P 〈0.01）.The apoptosis rates of the cell transfected with sh RNA-JNK/P38 group and miR-350 group were 16.36% and 8.39% respectively. And miR-350 inhibited the expression of endogenous p38 and JNK genes at the transcription level. Nevertheless this activity of miR-350 was significantly blocked by miR-350 antagomir.Conclusions miR-350 regulates proliferation and apoptosis of H9 c2 cell by targeting both MAPK14 and JNK gene.

关 键 词：心肌肥厚 miR-350 凋亡 

分 类 号：R541[医药卫生—心血管疾病]