沉默circMET靶向miR-30a-5p/ITGβ3抑制人直肠癌细胞株SW1463侵袭和迁移的作用研究  

Silencing circMET targets miR-30a-5p/ITGβ3 to inhibit the invasion and migration of human rectal cancer cell line SW1463

作　　者：李萌[1] 高志寒 于溯洋[1] LI Meng;GAO Zhi-han;YU Su-yang(Gastroenterology Department,Third Hospital of Hebei Medical University,Shijiazhuang 050051,China)

机构地区：[1]河北医科大学第三医院胃肠外科,石家庄050051

出　　处：《现代免疫学》2024年第4期346-352,共7页Current Immunology

基　　金：河北省2022年度医学科学研究课题计划项目(20220124)。

摘　　要：为探讨沉默环状核糖核酸MET(cyclic ribonucleic acid MET,circMET)靶向miR-30a-5p/整合素β3(integrinβ3,ITGβ3)抑制人直肠癌细胞侵袭和迁移的作用,选取人直肠癌细胞株SW1463进行相关研究。将SW1463细胞分为si-NC组、si-circMET组、miR-NC组、miR-30a-5p组、si-circMET+anti-miR-NC组、si-circMET+anti-miR-30a-5p组,每组6个复孔。通过Transwell小室试验检测细胞侵袭和迁移能力;qRT-PCR及Western blotting检测circMET、miR-30a-5p表达及ITGβ3、基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)、基质金属蛋白酶9(matrix metalloproteinase 9,MMP-9)表达水平;双荧光素酶报告基因试验分别验证circMET与miR-30a-5p、miR-30a-5p与ITGβ3的靶向关系。结果显示,与si-NC组比较,si-circMET组侵袭和迁移细胞数目,circMET表达及ITGβ3、MMP-2、MMP-9的mRNA和蛋白表达降低,miR-30a-5p表达升高(P<0.05);与miR-NC组比较,miR-30a-5p组侵袭和迁移细胞数目,circMET表达及ITGβ3、MMP-2、MMP-9的mRNA和蛋白表达降低,miR-30a-5p表达升高(P<0.05);与si-circMET+anti-miR-NC组比较,si-circMET+anti-miR-30a-5p组侵袭和迁移细胞数目,circMET表达及ITGβ3、MMP-2、MMP-9的mRNA和蛋白表达升高,miR-30a-5p表达降低(P<0.05);双荧光素酶报告基因试验结果证实,circMET与miR-30a-5p、miR-30a-5p与ITGβ3之间有靶向调控关系。综上所述,沉默circMET可抑制人直肠癌细胞株SW1463的侵袭和迁移,可能是通过靶向miR-30a-5p抑制ITGβ3、MMP-2、MMP-9的表达实现的。This study aims to investigate the effect of silencing cyclic ribonucleic acid MET(circMET)on inhibiting the invasion and migration of human rectal cancer cell by targeting miR-30a-5p/integrinβ3(ITGβ3).Human rectal cancer cell line SW1463 were divided into si-NC group,si-circMET group,miR-NC group,miR-30a-5p group,si-circMET+anti-miR-NC group,and si-circMET+anti-miR-30a-5p group,with 6 replicates in each group.Transwell assays were used to detect cell invasion and migration.The expressions of circMET,miR-30a-5p,ITGβ3,matrix metalloproteinase 2(MMP-2),and matrix metalloproteinase 9(MMP-9)were determined by qRT-PCR and Western blotting.Dual luciferase reporter gene assays were used to verify the targeting of circMET to miR-30a-5p and miR-30a-5p to ITGβ3,respectively.The results showed that compared to those of the si-NC group,the number of invasive and migrating cells,the expression of circMET,and the mRNA and protein expressions of ITGβ3,MMP-2,and MMP-9 in the si-circMET group were decreased,whereas the expression of miR-30a-5p was increased(P<0.05).Compared to those of the miR-NC group,the number of invasive and migrating cells,the expression of circMET,and the mRNA and protein levels of ITGβ3,MMP-2,and MMP-9 in the miR-30a-5p group were decreased,while the expression of miR-30a-5p was increased(P<0.05).Compared to those of the si-circMET+anti-miR-NC group,the number of invasive and migrating cells,the expression of circMET,and the mRNA and protein levels of ITGβ3,MMP-2,and MMP-9 in the si-circMET+anti-miR-30a-5p group were increased,while the expression of miR-30a-5p was decreased(P<0.05).In addition,Dual luciferase reporter gene assays confirmed the targeted regulatory relationship between circMET and miR-30a-5p as well as miR-30a-5p and ITGβ3.In summary,silencing circMET can inhibit the invasion and migration of human rectal cancer cell line SW1463,which may be achieved by targeting miR-30a-5p to inhibit the expressions of ITGβ3,MMP-2,and MMP-9.

关 键 词：环状核糖核酸MET 微小RNA-30a-5p 整合素Β3 直肠癌 侵袭 迁移 

分 类 号：R574.63[医药卫生—消化系统]