外泌体来源长链非编码RNA小核仁RNA宿主基因14对食管癌5-氟尿嘧啶耐药的影响  

Effect of exosome long non-coding RNA small nucleolus RNA host gene 14 on 5-fluorouracil resistance of esophageal cancer

作　　者：周昆[1] 赵志超 张芳宾[2] 闫焱[3] 李冰[1] Zhou Kun;Zhao Zhichao;Zhang Fangbin;Yan Yan;Li Bing(Department of Thoracic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Gastroenterology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Oncology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院胸外科,郑州450052 [2]郑州大学第一附属医院消化内科,郑州450052 [3]郑州大学第一附属医院肿瘤科,郑州450052

出　　处：《中华实验外科杂志》2023年第1期66-71,共6页Chinese Journal of Experimental Surgery

基　　金：河南省高等学校重点科研项目(20A320065);河南省医学科技攻关计划项目(201702010)。

摘　　要：目的探讨食管癌5-氟尿嘧啶(5-Fu)耐药细胞外泌体来源的长链非编码RNA(lncRNA)小核仁RNA宿主基因14(SNHG14)在食管癌耐药中的作用。方法实时荧光定量聚合酶链反应(RT-qPCR)检测郑州大学第一附属医院30例食管癌患者和30例健康对照血浆中lncRNA SNHG14的表达。浓度梯度法构建食管癌5-Fu耐药细胞3组EC9706/5-Fu和3组KYSE150/5-Fu,分离并收集各组食管癌细胞外泌体,RT-qPCR检测SNHG14的表达。分别将3组食管癌细胞EC9706和KYSE150与耐药细胞来源外泌体共培养,细胞计数试剂盒(CCK-8)检测5-Fu作用下细胞活性。敲低3组EC9706/5-Fu细胞中的SNHG14后分离外泌体,通过CCK-8、Transwell、Tunnel实验检测该外泌体对EC9706细胞药物敏感度、迁移和凋亡的影响。采用t检验或单因素方差分析(One-Way ANOVA)进行统计分析。结果食管癌患者血浆中lncRNA SNHG14的表达高于健康对照组(3.30±0.20比1.33±0.09,t=49.910,P<0.001),耐药食管癌细胞(EC9706/5-Fu和KYSE150/5-Fu)外泌体中lncRNA SNHG14的表达高于对照食管癌细胞(2.55±0.42比0.99±0.06,t=6.347;1.70±0.07比0.99±0.11,t=9.191,P<0.05)。在5-Fu作用下,EC9706/5-Fu-exo和KYSE150/5-Fu-exo组的细胞活性分别高于EC9706-exo和KYSE150-exo组[(165.23±5.97)%比(103.90±10.99)%,t=8.494;(139.35±3.82)%比(99.99±5.67)%,t=9.953,P<0.05]。EC9706/5-Fu-exo-si-SNHG14组细胞凋亡高于EC9706/5-Fu-exo组[(9.03±0.63)%比(5.53±0.73)%,F=40.290];细胞侵袭低于EC9706/5-Fu-exo组[(117.92±15.54)%比(339.52±27.43)%,F=152.000];5-Fu作用下细胞活性低于EC9706/5-Fu-exo组[(134.69±6.01)%比(160.38±4.69)%,F=64.090];细胞中JAK和p-STAT3的蛋白表达低于EC9706/5-Fu-exo组(0.86±0.07比1.87±0.10,F=363.100;0.96±0.08比1.83±0.06,F=388.700),差异有统计学意义(P<0.001)。结论食管癌耐药细胞外泌体来源lncRNA SNHG14能够通过激活JAK/STAT3通路提高食管癌对5-Fu的药物耐受。Objective To investigate the role of exosome long non-coding RNA(lncRNA)small nucleolus RNA host gene 14(SNHG14)derived from 5-fluorouracil(5-Fu)resistant cells in drug resistance of esophageal cancer.Methods The expression of lncRNA SNHG14 in plasma isolated from 30 esophageal cancer patients and 30 healthy controls were detected using real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).5-Fu drug resistant esophageal cancer cell lines,3 EC9706/5-Fu cell lines and 3 KYSE150/5-Fu cell lines,were constructed using concentration gradient method.Exosomes were isolated and collected from esophageal cancer cells in each group,and the expression of SNHG14 was measured by RT-qPCR.3 EC9706 or KYSE150 cells were cultured with exosomes isolated from drug-resistant esophageal cancer cells,and cell viability under 5-Fu treatment was detected by cell counting kit-8(CCK-8)assay.Exosomes were isolated from 3 SNHG14 knocked down EC9706/5-Fu cells,and the effects of the exosomes on drug sensitivity,migration and apoptosis of EC9706 cells were detected by CCK-8 assay,Transwell and Tunnel experiments,respectively.T test or one-way ANOVA was used for statistical analysis,and P<0.05 was considered statistically significant.Results The expression of lncRNA SNHG14 was significantly higher in plasma of esophageal cancer patients than in healthy controls(3.30±0.20 vs.1.33±0.09;t=49.910,P<0.001),and significantly higher in exosome derived from drug resistant esophageal cancer cells EC9706/5-Fu and KYSE150/5-Fu than in control esophageal cancer cells(2.55±0.42 vs.0.99±0.06,t=6.347;1.70±0.07 vs.0.99±0.11,t=9.191,P<0.05).Under 5-Fu treatment,cell viability was significantly higher in EC9706/5-Fu-exo and KYSE150/5-Fu-exo groups than EC9706-exo and KYSE150-exo groups,respectively[(165.23±5.97)%vs.(103.90±10.99)%,t=8.494;(139.35±3.82)%vs.(99.99±5.67)%,t=9.953,P<0.05].Cell apoptosis was higher[(9.03±0.63)%vs.(5.53±0.73)%,F=40.290],cell invasion was lower[(117.92±15.54)%vs.(339.52±27.43)%,F=152.000],cell viability

关 键 词：食管癌 外泌体 长链非编码RNA SNHG14 氟尿嘧啶 耐药 酪氨酸激酶/信号转导与转录激活因子3 

分 类 号：R735.1[医药卫生—肿瘤]