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(integrin β3,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.
Abstract
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/integrin β3(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.
基金项目
河北省2022年度医学科学研究课题计划项目(20220124)
NETL
NSTL
万方数据