微小核糖核酸-221通过调节Brahma相关基因-1促进结肠癌细胞侵袭转移的研究  

Promotion of microribonucleic acid-221 on invasion and metastasis of colon cancer cells through Brahma-related gene-1

作　　者：兰静芩 张根山 饶泽君 罗学来[1] 李海杰[1] Lan Jingqin;Zhang Genshan;Rao Zejun;Luo Xuelai;Li Haijie(GI Cancer Research Institute,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院胃肠肿瘤研究所,武汉430030

出　　处：《中华实验外科杂志》2023年第3期424-426,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81902851、82103283)。

摘　　要：目的探讨微小核糖核酸(RNA)-221(miR-221)对结肠癌细胞侵袭转移能力的影响及其分子机制。方法以人结肠癌SW48细胞作为研究对象,设置高表达对照组(miR-NC组),高表达miR-221组(miR-221组),低表达对照组(lnh-NC组),低表达miR-221组(lnh-221组),通过增加或降低miR-221表达,采用细胞迁移及侵袭试验(Transwell实验),检测miR-221对结肠癌细胞侵袭转移能力的影响;通过使用蛋白质印迹法(Western blot)检测miR-221对Brahma相关基因-1(BRG1)表达的影响。通过荧光素酶报告基因(Luciferase)实验,检测miR-221在SW48细胞中对BRG1转录活性的影响。通过在miR-221高表达的SW48细胞中高表达BRG1,检测BRG1在miR-221调控结肠癌细胞侵袭转移中的作用。组间比较采用t检验。结果在SW48细胞中,迁移和侵袭实验结果显示,miR-221组细胞的迁移和侵袭能力高于miR-NC组(迁移,miR-NC 110.30±7.79比miR-221193.00±4.73,t=9.071,P<0.01;侵袭,miR-NC 62.67±8.45比miR-221122.00±21.17,t=2.603,P<0.01);与lnh-NC组比较,lnh-221组细胞的迁移和侵袭能力低于lnh-NC组(迁移,lnh-NC 104.70±11.29比lnh-22173.67±6.12,t=2.410,P<0.01;侵袭,lnh-NC 55.67±8.11比lnh-22132.67±4.05,t=2.545,P<0.01)。在结肠癌LoVo细胞和SW48细胞中分别高表达和低表达miR-221,Western blot结果显示miR-221高表达可以使BRG1表达量降低,miR-221低表达可以使BRG1表达量升高。Luciferase实验中,miR-221高表达组BRG1 mRNA的转录活性低于miR-NC组(miR-NC 1.10±0.08比miR-2210.62±0.05,t=4.862,P<0.01),这提示miR-221可能直接与BRG1 mRNA 3’端非编码区(3’UTR)结合,发挥对BRG1的调控作用。在迁移和侵袭实验中,同时高表达miR-221和BRG1组(miR-221/Ad-BRG1组)细胞迁移和侵袭能力低于高表达miR-221组(miR-221/Ad-GFP组)(迁移,miR-221/Ad-GFP 163.30±9.39比miR-221/Ad-BRG127.33±2.33,t=14.063,P<0.01;侵袭,miR-221/Ad-GFP 75.33±8.19比miR-221/Ad-BRG111.67±1.33,t=7.674,P<0.01),这提示高表达BRG1可减弱miR-221促进结肠癌细胞Objective To investigate the effect of microribonucleic acid-221(miR-221)on the invasion and metastasis of colon cancer cells and its possible molecular mechanism.Methods The effect of miR-221 on the invasion and metastasis was detected by Transwell test and the effect of miR-221 on Brahma-related gene-1(BRG1)expression was detected by Western blotting in SW48.The effect of miR-221 on BRG1 transcription activity was detected by Luciferase experiment.The role of BRG1 in the regulation of invasion and metastasis by miR-221 was detected by high expression of BRG1 in SW48 cells with high expression of miR-221.T-test was used for comparisons between groups.Results In the migration and invasion experiment,the migration and invasion ability of cells in the miR-221 group was enhanced(migration:miR-NC 110.30±7.79 vs.miR-221193.00±4.73,t=9.071,P<0.01;invasion:miR-NC 62.67±8.45 vs.miR-221122.00±21.17,t=2.603,P<0.01).The migration and invasion ability of lnh-221 group decreased(migration:lnh-NC 104.70±11.29 vs.lnh-22173.67±6.12,t=2.410;invasion:lnh-NC 55.67±8.11 vs.lnh-22132.67±4.05,t=2.545,P<0.01).Western blotting showed that the high expression of miR-221 could reduce the expression of BRG1.In Luciferase experiment,the transcriptional activity of BRG1 mRNA in miR-221 overexpression group decreased(miR-NC 1.10±0.08 vs.miR-2210.62±0.05,t=4.862,P<0.01).This suggests that miR-221 may directly bind to BRG1 mRNA 3′UTR.In the migration and invasion experiment,the cell migration and invasion ability of the high expression miR-221 and BRG1 group(miR-221/Ad-BRG1 group)decreased(migration:miR-221/Ad-GFP 163.30±9.39 vs.miR-221/Ad-BRG127.33±2.33,t=14.063,P<0.01;invasion:miR-221/Ad-GFP 75.33±8.19 vs.miR-221/Ad-BRG111.67±1.33,t=7.674,P<0.01).This suggests that overexpression of BRG1 can reduce the ability of miR-221 to promote the invasion and metastasis.Conclusion MiR-221 can promote the invasion and metastasis of colon cancer cells,which may be partly dependent on BRG1.

关 键 词：结肠癌 微小核糖核酸 侵袭 转移 

分 类 号：R735.35[医药卫生—肿瘤]