抑制lncRNA TUG1靶向上调miR-26a介导VEGF/P38MAPK/Hsp27通路对结肠癌SW480细胞生物学行为影响  

Effect of inhibition of lncRNA TUG1 targeting up-regulation of miR-26a-mediated VEGF/P38MAPK/Hsp27 pathway on biological behavior of colon cancer SW480 cells

作　　者：李永坤[1] 贾廷印[1] 刘耿[1] LI Yong-Kun;JIA Ting-Yin;LIU Geng(General Surgery,First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,China)

机构地区：[1]南阳医学高等专科学校第一附属医院普通外科,南阳473000

出　　处：《中国免疫学杂志》2020年第20期2479-2484,共6页Chinese Journal of Immunology

基　　金：河南省科研计划基金资助项目(152102310025)。

摘　　要：目的:探究抑制干扰长链非编码RNA(lncRNA)TUG1靶向上调miR-26a介导VEGF/P38MAPK/Hsp27通路对结肠癌SW480细胞的生长、侵袭和EMT的影响。方法:sh-TUG1分别与miR-26a inhibitor和通路抑制剂SB203580单独或联合转染SW620细胞,以未经处理的SW480细胞为对照组。RT-PCR检测各组细胞TUG1及miR-26a表达,荧光素酶报告基因检测TUG1及miR-26a的靶向关系,BrdU法检测细胞增殖,Tanswell检测细胞侵袭,Western blot检测蛋白表达。将SW480细胞及转染sh-TUG1的SW480细胞分别皮下注射入裸鼠体内,分别设为对照组及sh-TUG1组,正常喂养30 d后取瘤、量体积,RT-PCR检测肿瘤组织TUG1及miR-26a表达,免疫组化检测肿瘤组织中Ki67、VEGF和Vimentin表达。结果:miR-26a是TUG1的靶向基因。相较于对照组,sh-TUG1组细胞增殖及侵袭数、Vimentin及N-cadherin蛋白表达明显降低,而E-cadherin蛋白表达明显升高(P<0.05);miR-26a inhibitor组细胞增殖及侵袭数、Vimentin及N-cadherin蛋白表达明显升高,而E-cadherin蛋白表达明显降低(P<0.05)。相较于miR-26a inhibitor组,sh-TUG1+inhibitor组细胞增殖及侵袭数、Vimentin及N-cadherin蛋白表达明显降低,而E-cadherin蛋白表达明显升高(P<0.05)。SB203580+LV-TUG1组细胞增殖及侵袭数、VEGF、P-P38/P38、P-Hsp27/Hsp27相对表达量高于SB203580组(P<0.05)。sh-TUG1组肿瘤体积,肿瘤组织中TUG1、Ki67、VEGF和Vimentin表达低于对照组,miR-26a表达高于对照组(P<0.05)。结论:抑制TUG1表达可靶向上调miR-26a水平,能通过抑制VEGF/P38MAPK/Hsp27通路抑制结肠癌SW480细胞的生长、运动和裸鼠成瘤。Objective:To explore inhibition of interfering with long-chain non-coding RNA(lncRNA)TUG1 targeting up-regulates effect of miR-26a-mediated VEGF/P38MAPK/Hsp27 pathway on growth,invasion,and EMT of colon carcinoma SW480 cells.Methods:sh-TUG1 was transfected into SW620 cells with miR-26a inhibitor and pathway inhibitor SB203580 separately or jointly,and untreated SW480 cells were used as control group.RT-PCR was used to detect expressions of TUG1 and miR-26a in each group,luciferase reporter gene was used to detect targeting relationship between TUG1 and miR-26a,cell proliferation was detected by BrdU method,cell invasion was detected by Tanswell and protein expression was detected by Western blot.SW480 cells and SW480 cells transfected with sh-TUG1 were injected subcutaneously into nude mice which were set as control group and sh-TUG1 group.After 30 d of normal feeding,tumors were taken to measure the volume.RT-PCR was used to detect expressions of TUG1 and miR-26a in tumor tissues,and immunohistochemistry was used to detect expressions of Ki67,VEGF and Vimentin in tumor tissues.Results:miR-26a was target gene of TUG1.Compared with control group,the cell proliferation and invasion quantities and protein expressions of Vimentin and N-cadherin in sh-TUG1 group were significantly decreased while protein expressions of E-cadherin was significantly increased(P<0.05).Cell proliferation and invasion quantities and protein expressions of Vimentin and N-cadher in miR-26a inhibitor group were significantly increased while protein expression of E-cadherin was significantly decreased(P<0.05).Compared with miR-26a inhibitor group,the cell proliferation and invasion quantities and protein expressions of Vimentin and N-cadherin in sh-TUG1+inhibitor group were significantly decreased while protein expression of E-cadherin was significantly increased(P<0.05).Cell proliferation and invasion quantities and relative expressions of VEGF,P-P38/P38 and P-Hsp27/Hsp27 in SB203580+LV-TUG1 group were higher than those in SB203580 group(P<0.

关 键 词：干扰长链非编码RNA TUG1 miR-26a 信号通路 结肠癌 SW480细胞 

分 类 号：R737.33[医药卫生—肿瘤]