PVT1通过调节miR-486-3p/MAPK14轴降低食管癌细胞的化学敏感性  

PVT1 reducing chemical sensitivity of esophageal cancer cells by regulating miR-486-3p/MAPK14 axis

作　　者：卢万里[1] 金哲[1] 段东奎[1] 王铮[1] 裴书飞 LU Wan-Li;JIN Zhe;DUAN Dong-Kui;WANG Zheng;PEI Shu-Fei(Department of Thoracic Surgery,Nanyang Central Hospital,Nanyang 473000,China)

机构地区：[1]南阳市中心医院胸外科,南阳473000 [2]南阳市中心医院消化科,南阳473000

出　　处：《中国免疫学杂志》2021年第24期2999-3004,共6页Chinese Journal of Immunology

基　　金：河南省医学科技攻关计划项目(LHGJ20200907)。

摘　　要：目的:探讨长链非编码RNA PVT1通过调节miR-486-3p/MAPK14轴降低食管癌细胞化学敏感性的相关机制。方法:qRT-PCR检测永生化人食管上皮细胞系和食管癌细胞系中PVT1、miR-486-3p、MAPK14的相对表达水平;生物信息学预测PVT1、miR-486-3p的靶向关系,并以荧光素酶报告实验验证靶向关系;构建PVT1、miR-486-3p、MAPK14过表达和PVT1敲除耐顺铂食管癌细胞系Eca-109/DDP,DDP处理后,比较各组Eca-109/DDP中的半抑制浓度(IC50)、细胞存活率、细胞凋亡率,并检测细胞凋亡蛋白(Bcl-2、Bax)、耐药蛋白(MRP1、P-gp)表达情况。结果:PVT1、MAPK14在食管癌细胞中高表达,miR-486-3p低表达(P<0.05);PVT1靶向抑制miR-486-3p表达,过表达PVT1后Eca-109/DDP细胞的IC50、细胞增殖率,MRP1、P-gp、Bcl-2表达水平显著升高(P<0.05),细胞凋亡率、Bax表达水平显著降低(P<0.05),过表达miR-486-3p能够恢复上述表型;敲除PVT1后Eca-109/DDP细胞的IC50、细胞增殖率、MAPK14表达水平显著降低(P<0.05),细胞凋亡率显著升高(P<0.05),过表达MAPK14后能够恢复上述表型。结论:PVT1可能通过靶向抑制miR-486-3p表达,促进MAPK14表达诱导耐药蛋白上调表达,降低食管癌细胞的化学敏感性。Objective:To explore related mechanism of long non-coding RNA PVT1 reducing chemical sensitivity of esophageal cancer cells by regulating miR-486-3p/MAPK14 axis.Methods:The qRT-PCR was applied to detect relative expression levels of PVT1,miR-486-3p and MAPK14 in immortalized human esophageal epithelial cell lines and esophageal cancer cell lines.The bioinformatics was applied to predict targeting relationship between PVT1 and miR-486-3p.And their targeting relationship was verified by luciferase report assay.The overexpression of PVT1,miR-486-3p and MAPK14,and PVT1-knockout cisplatin-resistant esophageal cancer cell line Eca-109/DDP were constructed.After DDP treatment,half inhibitory concentration(IC50)in Eca-109/DDP,cell survival rate and apoptosis rate were compared between all groups.The expression of apoptosis proteins(Bcl-2,Bax)and drug resistance proteins(MRP1,P-gp)was detected.Results:PVT1 and MAPK14 were highly expressed,while miR-486-3p was lowly expressed in esophageal cancer cells(P<0.05).PVT1 targetedly inhibited expression of miR-486-3p.After PVT1 overexpression,IC50 in Eca-109/DDP cells,cell proliferation rate,expression levels of MRP1,P-gp and Bcl-2 were significantly increased(P<0.05),while apoptosis rate and expression level of Bax were significantly decreased(P<0.05).The overexpression of miR-486-3p could restore the above phenotype.After knocking out PVT1,IC50 in Eca-109/DDP cells,cell proliferation rate and expression level of MAPK14 were significantly decreased(P<0.05),while apoptosis rate was significantly increased(P<0.05).The overexpression of MAPK14 could restore the above phenotype.Conclusion:PVT1 may promote expression of MAPK14,induce up-regulation of drug resistance proteins,and reduce chemical sensitivity of esophageal cancer cells by targetedly inhibiting expression of miR-486-3p.

关 键 词：食管鳞状细胞癌 PVT1 miR-486-3p MAPK14 顺铂敏感性 

分 类 号：Q291[生物学—细胞生物学]