摘要
目的探究miR-23a对染氟大鼠成骨肉瘤(UMR-106)细胞外调节蛋白激酶(ERKs)通路的影响。方法体外培养UMR-106细胞,分别转染miR-23a的激动剂(mimics)和抑制剂(inhibitor),再以0、80μmol/L NaF对细胞进行染毒,培养24 h后利用实时荧光定量PCR(qRT-PCR)检测ERKs通路关键因子细胞外信号调节激酶(ERK)、血清效应因子(SRF)、c-fos mRNA的表达水平;培养48 h后利用免疫蛋白印记法(Western blot)检测ERKs通路关键因子ERK、ETS样蛋白1(Elk-1)、SRF、c-fos蛋白的表达水平。结果与空白对照组比较,NaF组ERK、SRF、c-fos的mRNA和蛋白表达水平均升高,差异均有统计学意义(P<0.05)。转染miR-23a激动剂后,与miR-NC组比较,过表达组miR-23a、SRF mRNA表达水平上升(P<0.05),ERK、Elk-1、SRF蛋白表达水平上升(P<0.05),c-fos mRNA和蛋白表达变化不显著(P>0.05);转染mi R-23a抑制剂后,与Inhibitor NC组相比,抑制组miR-23a、ERK、SRF、c-fos mRNA表达水平降低(P<0.05),ERK、Elk-1、SRF、c-fos蛋白表达水平降低(P<0.05);转染miR-23a激动剂联合NaF作用下,与过表达组比较,过表达+NaF组ERK、SRF、cfos mRNA表达水平升高(P<0.05),ERK、Elk-1、c-fos蛋白表达水平升高(P<0.05);转染mi R-23a抑制剂联合NaF作用下,与抑制组比较,抑制+NaF组ERK、c-fos mRNA表达水平升高(P<0.05),ERK、SRF、c-fos蛋白表达水平上升(P<0.05)。结论miR-23a能激活染氟UMR-106细胞ERKs通路途径。
Objective To understand the effects of miR-23 a on the extracellular regulatory protein kinases(ERKs)pathway in rat osteosarcoma(UMR-106)exposed to fluoride.Methods UMR-106 cells were cultured in vitro,and miR-23 a agonists(mimics)and inhibitors were transfected respectively.The cells were treated with NaF at the doses of 0 and 80μmol/L respectively.After 24 hours-treatment,the mRNA expression levels of critical factors in miR-23 a and ERKs pathway,extracellular signal-regulated kinase(ERK),SRF and c-fos,were detected by qRT-PCR;After 48 hours-treatment,the expression levels of various factors protein in ERKs pathway were detected by Western blot.Results Compared with the control group,the mRNA and protein expression levels of ERK,SRF and c-fos significantly increased(P<0.05).After transfection of miR-23 a agonist,compared with miR-NC group,the expression level of SRF,miR-23 a mRNA in over expression group increased(P<0.05),and the expression levels of ERK,Elk-1 and SRF protein increased significantly(P<0.05),and no significant changes were observed in c-fos mRNA and protein expression(P>0.05);After transfection of miR-23 a inhibitor,compared with miR-NC group,the expression of miR-23 a,ERK,SRF and c-fos mRNA in the inhibition group was decreased(P<0.05),and the protein expressions of ERK,Elk-1,SRF and c-fos were decreased in the inhibition group(P<0.05).The expression levels of ERK,SRF and c-fos mRNA,the expression levels of ERK,Elk-1,c-fos protein increased in miR-23 a over expression+NaF group compared with those in over expression group(P<0.05).Compared with the inhibitory group,the expression of ERK and c-fos mRNA,the expression levels of ERK,SRF and c-fos protein increased in the miR-23 a inhibitory expression+NaF group(P<0.05).Conclusion miR-23 a can activate the ERKs pathway in UMR-106 cells exposed to fluoride.
作者
杨璐珲
卢明丽
陈宣好
王楠兰
韦艳
YANG Lu-hui;LU Ming-li;CHEN Xuan-hao;WAN Nan-lan;WEI Yan(School of Public Health,the Key Laboratory of Environmental Pollution Monitoring and Disease Control,Ministry of Education,Guizhou Medical University,Guiyang,Guizhou 550025,China;不详)
出处
《环境与健康杂志》
CAS
北大核心
2020年第3期189-194,共6页
Journal of Environment and Health
基金
国家自然科学基金(81560515)
