干扰长链非编码RNAMALAT1对结肠癌SW480细胞生长、侵袭和裸鼠成瘤的影响  

Effects of interfering with long non-coding RNAMALAT1 on colon cancer SW480 cell growth and invasion,and tumorigenicity in nude mice

作　　者：裴正浩 王耿泽[1] 夏西超 张虎 王钧[1] 郝阳 PEI Zhenghao*;WANG Gengze;XIA Xichao;ZHANG Hu;WANG Jun;HAO Yang(Department of Gastrointestinal Surgery,Nanyang Central Hospital,Nanyang 473000,China;Medical School of Pingdingshan University,Pingdingshan 467000,China)

机构地区：[1]南阳市中心医院胃肠外科,河南南阳473000 [2]平顶山学院医学院,河南平顶山467000

出　　处：《广东药科大学学报》2022年第5期83-89,共7页Journal of Guangdong Pharmaceutical University

基　　金：河南省自然科学基金项目(182300410123)。

摘　　要：目的探讨干扰肺腺癌转移相关转录本1(MALAT1)对结肠癌SW480细胞生长、侵袭及裸鼠成瘤的影响及其作用机制。方法构建shRNA-MALAT1质粒载体,并与miR-145 inhibitor单独或联合转染人结肠癌SW480细胞,以转染空载体的细胞作为Control组。采用双荧光素酶报告基因实验验证MALAT1与miR-145靶向关系;qRT-PCR检测转染效率;分别使用Brdu法、Transwell实验检测SW480细胞增殖、侵袭;Western blot及免疫荧光检测细胞侵袭相关蛋白表达。取10只裸鼠,分别于皮下注射转染空载体(Control组)和shRNA-MALAT1(shRNA-MALAT1组)的SW480细胞构建结直肠癌移植瘤模型,测量肿瘤体积变化并于30 d后取瘤,采用免疫组化法检测肿瘤组织中Ki67、VEGF表达。结果共转染miR-145 mimic和野生型MALAT1报告基因载体后,SW480细胞的荧光素酶活性显著降低(P<0.05)。与Control组比较,shRNA-MALAT1组Brdu阳性细胞数、侵袭细胞数、N-cadherin、Vimentin、JAK2、p-STAT3/STAT3均显著降低(P<0.05),E-cadherin蛋白表达显著增加(P<0.05);miR-145 inhibitor组Brdu阳性细胞数、侵袭细胞数、N-cadherin、Vimentin、JAK2、p-STAT3/STAT3均显著增加(P<0.05),E-cadherin蛋白表达显著降低(P<0.05)。与miR-145 inhibitor组比较,shRNA-MALAT1+miR-145 inhibitor组Brdu阳性细胞数、侵袭细胞数、N-cadherin、Vimentin、JAK2、p-STAT3/STAT3均显著降低(P<0.05),E-cadherin蛋白表达显著增加(P<0.05)。体内实验中,与Control组比较,shRNA-MALAT1组裸鼠成瘤体积显著降低(P<0.05),肿瘤组织中Ki67、VEGF蛋白表达显著降低(P<0.05)。结论敲低MALAT1可通过负调控miR-145抑制结直肠癌SW480细胞的增殖、侵袭及体内成瘤能力。Objective To explore the effect and mechanism of interfering with metastasis-associated lung adenocarcinoma transcript 1(MALAT1)on colon cancer SW480 cell growth and invasion,and tumorigenicity in nude mice.Methods The shRNA-MALAT1 plasmid vector was constructed and transfected into SW480 cells alone or in combination with miR-145 inhibitor.Cells transfected with empty vector were used as the control group.Dual luciferase reporter gene experiment was used to verify the targeting relationship between MALAT1 and miR-145.The transfection efficiency was detected by qRT-PCR.The proliferation and invasion of SW480 cells were detected by BrdU assay and Transwell assay,respectively.The expression of invasion-related proteins was detected by western blot and immunofluorescence.Ten nude mice were injected subcutaneously with SW480 cells transfected with empty vector(control group)and shRNA-MALAT1(shRNA-MALAT1 group)to construct colorectal cancer transplanted tumor model.The tumor volume was measured and the tumor was removed 30 days later.The expression of Ki67 and VEGF in tumor tissue was detected by immunohistochemistry.Results After cotransfecting miR-145 mimic and wild-type MALAT1 reporter vectors,the luciferase activity of SW480 cells was significantly decreased(P<0.05).Compared with the control group,the number of Brdu positive cells and invasion cells,and the expression of N-cadherin,vimentin,JAK2 and p-STAT3/STAT3 were significantly decreased in shRNA-MALAT1 group(P<0.05),while the expression of E-cadherin was significantly increased(P<0.05).Compared with the control group,the number of Brdu positive cells and invasion cells,and the expression of N-cadherin,vimentin,JAK2 and p-STAT3/STAT3 were markedly increased in miR-145 inhibitor group(P<0.05),while the expression of E-cadherin was decreased(P<0.05).Compared with miR-145 inhibitor group,the number of Brdu positive cells and invasion cells,and the expression of N-cadherin,vimentin,JAK2 and p-STAT3/STAT3 were obviously lowered in shRNA-MALAT1+miR-145 inhibitor gro

关 键 词：结直肠癌 肺腺癌转移相关转录本1 MIR-145 增殖 侵袭 JAK2/STAT3 

分 类 号：R735.3[医药卫生—肿瘤]