miR-146a通过靶向CCNJ调控鼻咽癌细胞HNE-1/DDP对顺铂敏感性的影响

Effect of miR-146a on the sensitivity of nasopharyngeal carcinoma cells HNE-1/DDP to cisplatin by targeting CCNJ

目的探讨miR-146a对鼻咽癌耐顺铂细胞HNE-1/DDP顺铂敏感性的作用及可能机制。方法实验分空白组、阴性对照组及mimics-miR-146a组。MTT检测HNE-1/DDP细胞在不同方法处理后的增殖抑制作用;Annexin V-FITC/PI双染法检测细胞凋亡情况;Western blot检测周期蛋白CCNJ、MRP1、P-gp的相对表达水平;实时荧光定量PCR(RT-qPCR)检测CCNJ mRNA的相对表达水平。结果与其他两组相比,mimics-miR-146a组存活率降低,凋亡率升高,MRP1、P-gp、CCNJ蛋白表达量降低,CCNJ mRNA的相对表达量降低,差异均有统计学意义(P<0.05)。结论miR-146a可增加HNE-1/DDP对DDP的敏感性,机制可能是miR-146a抑制CCNJ的表达而调控下游蛋白MRP1、P-gp,降低细胞耐药性。

Objective To investigate the effect of miR-146a on cisplatin sensitivity of nasopharyngeal carcinoma HNE-1/DDP cells and its possible mechanism.Methods The experiment was divided into three groups:a blank control group,a negative control group and a mimics-miR-146a group.MTT assay was used to detect the proliferation inhibition of cells after different treatments.Annexin V-FITC/PI assay was used to detect apoptosis.Western blot was used to detect the relative expression levels of cyclin CCNJ,MRP1,P-gp.RT-qPCR was used to detect the relative expression level of CCNJ mRNA.Results Compared with other two groups,the survival rate,the relative protein expression levels of MRP1,P-gp,CCNJ and the relative expression level of CCNJ mRNA of mimics-miR-146a reduced,while the apoptosis rate increased.There were statistical differences in the survival rate,the relative protein expression levels of MRP1,P-gp,CCNJ,the relative expression level of CCNJ mRNA and the apoptosis rate among the three groups(P<0.05).Conclusions miR-146a can increase the sensitivity of HNE-1/DDP to DDP which mechanism may be through miR-146a inhibiting the expression of CCNJ to regulate downstream proteins MRP1,P-gp,and then to reduce cell resistance.