长链非编码RNA-ROR介导上皮-间质转化对鼻咽癌细胞放疗抵抗作用的体外研究  

Effect of long non-coding RNA-ROR mediating epithelial-mesenchymal transformation on radiotherapy resistance of nasopharyngeal carcinoma cells in vitro

作　　者：薛晓成[1] 张雪[2] 黄水仙 张燚[1] 鲁丹 陈晓平[1] XUE Xiaocheng;ZHANG Xue;HUANG Shuixian;ZHANG Yi;LU Dan;CHEN Xiaoping(Department of Otorhinolaryngology Head and Neck Surgery,Gongli Hospital,Naval Medical University(Second Military Medical University),Shanghai 200135,China;Key Laboratory of Artificial Intelligence for Inflammation and Chronic Disease Management,Shanghai Municipal Health Commission,Gongli Hospital,Naval Medical University(Second Military Medical University),Shanghai 200135,China)

机构地区：[1]海军军医大学(第二军医大学)附属公利医院耳鼻咽喉头颈外科,上海200135 [2]海军军医大学(第二军医大学)附属公利医院、上海市卫生健康委员会炎症与慢病管理人工智能重点实验室,上海200135

出　　处：《海军军医大学学报》2024年第10期1218-1225,共8页Academic Journal of Naval Medical University

基　　金：上海市浦东新区卫生系统优秀青年医学人才培养计划(PWRq-2020-63);上海市卫生健康委员会青年项目(20204Y0144);上海市浦东新区重点亚专科项目(PWZy2020-06);上海市浦东新区临床特色专科项目(PWYts2021-15)。

摘　　要：目的 探讨lncRNA-ROR介导上皮-间质转化在鼻咽癌细胞放疗抵抗中的作用。方法 将鼻咽癌细胞CNE2分为空白组、阴性对照组、lncRNA-ROR沉默组,进行相应的处理。将CNE2细胞分为空白组、放疗组、放疗+阴性对照组、放疗+lncRNA-ROR过表达组(放疗处理为6 Gy射线照射24 h),进行相应的处理。用CCK-8法检测CNE2增殖能力,通过细胞划痕实验和Transwell 细胞迁移实验检测细胞迁移能力,用流式细胞术检测细胞凋亡的情况,用蛋白印迹法检测凋亡相关蛋白和上皮-间质转化相关蛋白的表达。结果 与空白组、阴性对照组相比,抑制lncRNA-ROR表达48、72 h后鼻咽癌细胞CNE2的增殖能力均减弱(均P<0.05)。抑制lncRNA-ROR表达后鼻咽癌细胞CNE2的迁移率低于阴性对照组(P<0.05),而放疗+lncRNA-ROR过表达组CNE2细胞的迁移能力高于放疗组与放疗+阴性对照组(均P<0.05)。与放疗组、放疗+阴性对照组相比,放疗+lncRNA-ROR过表达组CNE2细胞的凋亡率均降低(均P<0.05)。抑制lncRNA-ROR后,活化的caspase 3、caspase 9蛋白表达均较空白组和阴性对照组升高(均P<0.05);而放疗+lncRNA-ROR过表达组活化的caspase 3、caspase 9蛋白表达均较放疗组和放疗+阴性对照组下降(均P<0.05)。抑制lncRNA-ROR可导致上皮标志蛋白(E-钙黏蛋白、β-联蛋白)表达升高,间质标志蛋白(N-钙黏蛋白、波形蛋白)表达下降(均P<0.05);而与放疗组和放疗+阴性对照组相比,放疗+lncRNA-ROR过表达组CNE2细胞的上皮标志蛋白表达下降、间质标志蛋白表达升高(均P<0.05)。结论 lncRNA-ROR可通过调控鼻咽癌细胞增殖、迁移、凋亡及上皮-间质转化影响其放疗抵抗,是逆转鼻咽癌细胞放疗抵抗的潜在靶点。Objective To investigate the role of long non-coding RNA(lncRNA)-ROR in mediating epithelial-mesenchymal transformation(EMT)and its impact on radiotherapy resistance in nasopharyngeal carcinoma cells.Methods Nasopharyngeal carcinoma cells CNE2 were divided into blank group,negative control(NC)group and lncRNA-ROR siliencing group;or were divided into blank group,radiotherapy group,radiotherapy+NC group,and radiotherapy+lncRNA-ROR overexpression group(radiotherapy treated with 6 Gy radiation for 24 h).The CNE2 proliferation was detected by cell counting kit 8 method.The cell migration was detected by cell scratch test and Transwell cell migration test.The apoptosis ratio was detected byflow cytometry,and the apoptosis-related proteins and epithelial-mesenchymal transition proteins were detected by Western blotting.Results Compared with the blank group and NC group,the proliferation ability of nasopharyngeal carcinoma cells CNE2 was decreased after inhibition of lncRNA-ROR expression for 48 and 72 h(all P<0.05).The mobility of CNE2 cells after lncRNA-ROR expression inhibition was lower than that in the NC group(P<0.05).The migration ability of CNE2 cells in the radiotherapy+lncRNA-ROR overexpression group was higher than that in the radiotherapy group and radiotherapy+NC group(both P<0.05).Compared with the radiotherapy group and radiotherapy+NC group,the apoptosis rates of CNE2 cells in the radiotherapy+lncRNA-ROR overexpression group was decreased(both P<0.05).After lncRNA-ROR inhibition,the expression of activated caspase 3 and caspase 9 proteins was increased(both P<0.05),while the expression of activated caspase 3 and caspase 9 proteins was decreased in the radiotherapy+overexpressed lncRNA-ROR group(both P<0.05).Inhibition of lncRNA-ROR increased the expression of epithelial marker proteins(E-cadherin,β-catenin),and decreased the expression of interstitial marker proteins(N-cadherin,vimentin).The epithelial marker protein expression was decreased and interstitial marker protein expression was increased in C

关 键 词：鼻咽肿瘤 放疗抵抗 上皮-间质转化 长链非编码RNA-ROR 

分 类 号：R739.63[医药卫生—肿瘤]