微小RNA-135b-5p对食管鳞癌增殖、迁移、侵袭的作用及机制  

Effect of microRNA-135b-5p on proliferation,migration and invasion of esophageal squamous cell carcinoma and underlying mechanism

作　　者：晁策 马超 张友浦 刘杨[1] 吕浩 狄冬梅[1] 张晓膺[1] Chao Ce;Ma Chao;Zhang Youpu;Liu Yang;Lyu Hao;Di Dongmei;Zhang Xiaoying(Department of Cardiothoracic Surgery,the Third Affiliated Hospital of Soochow University,Changzhou 213003,China)

机构地区：[1]苏州大学附属第三医院心胸外科,常州213003

出　　处：《中华实验外科杂志》2022年第8期1491-1493,共3页Chinese Journal of Experimental Surgery

基　　金：江苏省自然科学基金面上项目(BK20191158);常州市科技局社会发展支撑项目(CE20205039);常州市科技局应用基础研究(CJ20200104);常州市卫健委青苗人才项目(CZQM2020004)。

摘　　要：目的探讨微小RNA(miR)-135b-5p在食管鳞癌发生发展中的调节作用和机制。方法通过收集食管鳞癌患者的组织标本28例, 检测miR-135b-5p在食管鳞癌组织中的表达水平, 分析比较高表达与低表达组的差异, 采用细胞计数试剂盒(CCK-8)、划痕实验、Transwell实验及蛋白质印迹法(Western blot)实验探究miR-135b-5p在食管鳞癌细胞中的调节作用, 利用生物信息学技术从数据库中寻找miR-135b-5p的下游信号通路。组间比较采用T检验或卡方检验。结果 miR-135b-5p在食管鳞癌患者中低表达(0.004比0.014, Z=-2.89, P<0.01)且与T分期相关(χ^(2)=5.82, P<0.05)。此外, 划痕实验发现24 h后对照组划痕空白面积低于高表达组(ECA109:0.22±0.02比0.26±0.01, t=2.21, P<0.05;KYSE150:0.33±0.01比0.54±0.02, t=9.77, P<0.01), Transwell实验说明24 h后高表达组细胞迁移数量低于对照组(ECA109:47.17±4.98比136.00±7.78, t=9.62, P<0.01;KYSE150:25.83±2.10比153.00±7.49, t=16.34, P<0.01), 48 h后高表达组细胞侵袭数量低于对照组(ECA109:171.00±24.02比382.30±15.34, t=7.42, P<0.01;KYSE150:244.00±27.33比493.70±53.23, t=4.17, P<0.01), 高表达组p-smad2/3蛋白表达低于对照组(ECA109:0.82±0.02比1.10±0.03, t=7.70, P<0.01;KYSE150:0.28±0.02比0.73±0.02, t=14.12, P<0.01)。结论 miR-135b-5p可能通过下调smad2/3磷酸化水平抑制食管鳞癌细胞的迁移与侵袭。Objective To explore the effect and regulatory mechanism of microRNA(miR)-135b-5p in esophageal squamous cell carcinoma(ESCC).Methods Through collecting the 28 cases of clinical ESCC tissues,we analyzed the relative expression of miR-135b-5p by real time-polymerase chain reaction(RT-PCR)in ESCC.Meanwhile,cell counting kit-8(CCK-8),the wound healing,Transwell,Western blotting,and bioinformatics were used to evaluate the regulatory effect and detailed mechanism of miR-135b-5p in ESCC cells.The student’s test or chi-square test was used for between-group analysis.Results MiR-135b-5p expression was lower in ESCC tissue than match normal esophageal epithelial tissue(0.004 vs.0.014,Z=-2.89,P<0.01).The low expression of miR-135b-5p may be correlated with the T stage in ESCC patients(χ^(2)=5.82,P<0.05).Moreover,the scratch blank area was lower in the control group than in the high expression group after 24 h(ECA109:0.22±0.02 vs.0.26±0.01,t=2.21,P<0.05;KYSE150:0.33±0.01 vs.0.54±0.02,t=9.77,P<0.01).Compared with the control group,the number of migrating cells(ECA109:47.17±4.98 vs.136.00±7.78,t=9.62,P<0.01;KYSE150:25.83±2.10 vs.153.00±7.49,t=16.34,P<0.01)and invasive cells(ECA109:171.00±24.02 vs.382.30±15.34,t=7.42,P<0.01;KYSE150:244.00±27.33 vs.493.70±53.23,t=4.17,P<0.01)in the high expression group was significantly decreased.Compared with the control group,the p-smad2/3 expression level in high expression group was significantly decreased(ECA109:0.82±0.02 vs.1.10±0.03,t=7.70,P<0.01;KYSE150:0.28±0.02 vs.0.73±0.02,t=14.12,P<0.01).Conclusion MiR-135b-5p may down-regulate the phosphorylation expression of smad2/3 to suppress the migration and invasion of ESCC.

关 键 词：食管鳞癌 微小RNA 迁移 侵袭 

分 类 号：R735.1[医药卫生—肿瘤]