节点文献

microRNA-96-5p对染氟成骨肉瘤细胞c-Jun氨基末端激酶通路的影响

Effects of miR-96-5p on JNK pathway in UMR-106 cells treated with sodium fluoride

  • 推荐 CAJ下载
  • PDF下载
  • 不支持迅雷等下载工具,请取消加速工具后下载。

【作者】 陈宣好杨璐珲代佑罡吴彦秋王楠兰韦艳

【Author】 CHEN Xuan-hao;YANG Lu-hui;DAI You-gang;WU Yan-qiu;WANG Nan-lan;WEI Yan;Department of Occupational Health and Environmental Hygiene,School of Public Health,Guizhou Medical University;

【通讯作者】 韦艳;

【机构】 贵州医科大学公共卫生学院职业卫生与环境卫生学系贵州医科大学环境污染与疾病监控教育部重点实验室贵州医科大学公共卫生学院营养与食品安全学教研室

【摘要】 目的探讨miR-96-5p对染氟大鼠成骨肉瘤(UMR-106)细胞c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)通路的影响。方法将处于对数生长期的UMR-106细胞分别转染mi R-96-5p反义寡核苷酸(anti-miRNA oligonucleotide,AMO)、过表达质粒以达到抑制和过表达作用,再以0(对照)、2 000μmol/L NaF对细胞进行染毒,培养24 h后通过实时荧光定量PCR检测miR-96-5p及JNK通路上各因子基因的变化;培养48 h后,采用蛋白免疫印迹法检测JNK通路上各因子蛋白的变化。结果与无氟对照组相比,氟染毒组UMR-106细胞mi R-96-5p、天冬氨酸蛋白水解酶3(cysteinyl aspartate specific proteinase 3,caspase-3)mRNA表达水平升高(P<0.05),cAMP直接活化交换蛋白(exchange protein directly activated by cAMP,Epac)mRNA表达水平下降,差异均有统计学意义(P<0.05);转染mi R-96-5p质粒后,与质粒对照组相比,质粒+NaF组UMR-106细胞miR-96-5p mRNA表达水平升高,差异有统计学意义(P<0.05);转染AMO后,与AMO对照组相比,AMO+NaF组UMR-106细胞mi R-96-5p mRNA表达水平升高,Epac和caspase-3 mRNA表达水平下降,差异均有统计学意义(P<0.05)。与无氟对照组相比,氟染毒组UMR-106细胞腺苷酸环化酶6(Adenylate Cyclase 6,AC6)、Epac、p-JNK蛋白表达水平均下降,差异均有统计学意义(P<0.05);与质粒对照组相比,质粒+NaF组UMR-106细胞AC6、Epac、天冬氨酸蛋白水解酶9(cysteinyl aspartate specific proteinase 9,caspase-9)蛋白表达水平均下降,而p-JNK、磷酸化Jun癌基因(Jun oncogene,c-Jun)、细胞色素C(cytochrome C,cyt C)和非活化caspase-3蛋白表达水平升高,差异均有统计学意义(P<0.05);转染AMO后,与AMO对照组相比,AMO+NaF组UMR-106细胞AC6、Epac、caspase-9、p-JNK蛋白表达均下降,p-c-Jun、cyt C和非活化caspase-3蛋白表达升高,差异均有统计学意义(P<0.05)。结论 mi R-96-5p可能促进染氟UMR-106细胞JNK通路磷酸化,从而介导细胞凋亡。

【Abstract】 Objective To explore the effects of miR-96-5 p on JNK pathway in UMR-106 cells treated with NaF. Methods UMR-106 cells,transfected with miR-96-5 p vector and anti-miRNA oligonucleotide(AMO),exposed to 0 and 2 000 μmol/L NaF respectively,were determined for the gene expression of miR-96-5 p and JNK pathway by real-time quantitative PCR based on 24-hour culture,as well as the expression of proteins in JNK pathway by Western blot based on 48-hour culture.Results Compared with the control group,mi R-96-5 p and caspase-3 gene expression increased and exchange protein directly activated by cAMP(Epac) decreased significantly(P<0.05) in Naf exposure group. Compared with miR-96-5 p vector control group,miR-96-5 p was up-regulated(P <0.05),other factors’ m RNA did not change in mi R-96-5 p vector +NaF group.Compared with AMO control group,mi R-96-5 p gene was up-regulated(P<0.05),Epac and caspase-3 were down-regulated(P<0.05) in AMO+NaF group. Compared with the control group,the expression of AC6,Epac and p-JNK decreased(P<0.05) in Naf exposure group. Significantly lower expression of AC6,Epac and caspase-9,higher expression of p-JNK,p-c-Jun,cyt C,non-activated caspase-3 were found in miR-96-5 p vector+NaF group compared with those in mi R-96-5 p vector control group(P<0.05). Significantly lower expression of AC6,Epac,caspase-9 and p-JNK,higher expression of p-Jun,cyt C,non-activated caspase-3,were found in AMO+NaF group compared with those in AMO control group(P<0.05). Conclusion mi R-96-5 p may promote apoptosis via phosphorylating JNK pathway in UMR-106 cells exposed to NaF.

【基金】 国家自然科学基金(81560515)
  • 【文献出处】 环境与健康杂志 ,Journal of Environment and Health , 编辑部邮箱 ,2018年10期
  • 【分类号】R599.1
  • 【下载频次】24
节点文献中: 

本文链接的文献网络图示:

本文的引文网络