长链非编码RNA淋巴细胞白血病缺失基因2/微小RNA-30c-5p/丝裂原活化蛋白激酶1轴在食管癌细胞紫杉醇耐药中的作用机制实验研究  

Mechanism of lncRNA DLEU2/miR-30c-5p/MAPK1 axis in paclitaxel resistance in esophageal cancer cells

作　　者：符梦婕 张剑[1] 李秋忆 王晨[1] 石禹 周玲 FU Mengjie;ZHANG Jian;LI Qiuyi;WANG Chen;SHI Yu;ZHOU Ling(Department of Clinical Pharmacy,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院临床药学部,四川成都610041

出　　处：《陕西医学杂志》2024年第9期1155-1160,共6页Shaanxi Medical Journal

基　　金：国家自然科学基金青年科学基金资助项目(82304399)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)淋巴细胞白血病缺失基因2(DLEU2)/微小RNA(miR)-30c-5p/丝裂原活化蛋白激酶1(MAPK1)轴在食管癌细胞紫杉醇(PTX)耐药中的作用机制。方法:构建EC109/PTX细胞,检测食管癌组织和癌旁正常组织,以及食管癌细胞EC109和KYSE150细胞、食管上皮细胞SHEE和EC109/PTX细胞DLEU2、miR-30c-5p、MAPK1 mRNA表达。取生长良好的EC109/PTX细胞,分为沉默(siRNA)DLEU2组、阴性对照(siRNA NC)组、siRNA DLEU2+miR-30c-5p抑制剂(inhibitor)组、siRNA DLEU2+inhibitor NC组、siRNA DLEU2+过表达(pcDNA)MAPK1组、siRNA DLEU2+pcDNA NC组和空白组。验证miR-30c-5p与DLEU2、MAPK1的靶向关系。检测各组EC109/PTX细胞DLEU2、miR-30c-5p、MAPK1 mRNA表达水平。检测EC109/PTX细胞活性、凋亡情况及对PTX的耐药性。检测谷胱甘肽S-转移酶π(GST-π)、MAPK1、P-蛋白(P-gp)蛋白表达水平。结果:食管癌组织和EC109/PTX细胞miR-30c-5p表达降低,DLEU2、MAPK1 mRNA表达增加(均P<0.05)。DLEU2靶向调控miR-30c-5p,miR-30c-5p靶向调控MAPK1。siRNA DLEU2组miR-30c-5p表达和细胞凋亡率较siRNA NC组、空白组增加,DLEU2和MAPK1 mRNA表达、细胞增殖率、药物半数抑制浓度(IC 50)以及GST-π、MAPK1、P-gp蛋白表达降低(均P<0.05)。siRNA DLEU2+miR-30c-5p inhibitor组miR-30c-5p表达和细胞凋亡率较siRNA DLEU2+inhibitor NC组降低,MAPK1 mRNA表达、细胞增殖率、IC 50以及GST-π、MAPK1、P-gp蛋白表达增加(均P<0.05)。siRNA DLEU2+pcDNA MAPK1组细胞凋亡率较siRNA DLEU2+pcDNA NC组降低,MAPK1 mRNA表达、细胞增殖率、IC 50以及GST-π、MAPK1、P-gp蛋白表达增加(均P<0.05)。结论:干扰lncRNA DLEU2可减弱EC109/PTX细胞对PTX的耐药性,可能与调控miR-30c-5p/MAPK1轴有关。Objective:To investigate the mechanism of long non-coding RNA(lncRNA)deleted in lymphocytic leukemia 2(DLEU2)/microRNA(miR)-30c-5p/mitogen-activated protein kinase 1(MAPK1)axis in paclitaxel(PTX)resistance in esophageal cancer cells.Methods:EC109/PTX cells were constructed and the DLEU2,miR-30c-5p,and MAPK1 mRNA expressions in esophageal cancer tissue,normal tissue,esophageal cancer cells EC109 and KYSE150 cells,esophageal epithelial cells SHEE,and EC109/PTX cells were detected.The well growing EC109/PTX cells were taken and were separated into silenced(siRNA)DLEU2 group,negative control(siRNA NC)group,siRNA DLEU2+miR-30c-5p inhibitor group,siRNA DLEU2+inhibitor NC group,siRNA DLEU2+overexpression(pcDNA)MAPK1 group,siRNA DLEU2+pcDNA NC group and blank group,the targeting relationship between miR-30c-5p and DLEU2 and MAPK1 was verified.The expression levels of DLEU2,miR-30c-5p,MAPK1 mRNA,EC109/PTX cell activity,apoptosis,and resistance to PTX,and the expression levels of GST-π,MAPK1 and P-gp proteins in EC109/PTX cells in each group were detected.Results:The expression of miR-30c-5p decreased in esophageal cancer tissue and EC109/PTX cells,while the expressions of DLEU2 and MAPK1 mRNA increased(all P<0.05).DLEU2 targeted regulation of miR-30c-5p,and miR-30c-5p targeted regulation of MAPK1.The siRNA DLEU2 group showed a obvious increase in miR-30c-5p expression and apoptosis rate compared to the siRNA NC group and blank group,while the DLEU2 and MAPK1 mRNA expression,proliferation rate,IC 50 value,and GST-π,MAPK1,and P-gp protein expressions were obviously reduced(all P<0.05).The siRNA DLEU2+miR-30c-5p inhibitor group showed a obvious decrease in miR-30c-5p expression and apoptosis rate compared to the siRNA DLEU2+inhibitor NC group,while the MAPK1 mRNA expression,proliferation rate,IC 50 value,and GST-π,MAPK1,and P-gp protein expressions were obviously increased(all P<0.05).The apoptosis rate of the siRNA DLEU2+pcDNA MAPK1 group was obviously lower than that of the siRNA DLEU2+pcDNA NC group,while the MAPK1 mRN

关 键 词：食管癌 长链非编码RNA DLEU2 微小RNA-30c-5p 丝裂原活化蛋白激酶1 紫杉醇 耐药性 

分 类 号：R73-3[医药卫生—肿瘤]