摘要
目的:探讨CircDONSON对肝癌Huh-7细胞增殖、迁移及侵袭的影响及其可能的作用机制。方法:qRT-PCR法检测肝癌组织中CircDONSON、miR-4458表达量;肝癌细胞Huh-7分为si-NC组、si-CircDONSON组、miR-NC组、miR-4458组、si-CircDONSON+anti-miR-NC组、si-CircDONSON+anti-miR-4458组;CCK-8法、平板克隆形成实验、划痕实验与Transwell实验分别检测细胞增殖、克隆形成、迁移及侵袭情况;双荧光素酶报告实验检测miR-4458与CircDONSON靶向的关系;Western blot检测MMP-2、MMP-9蛋白表达量。结果:肝癌组织中CircDONSON表达量较癌旁组织升高(4.82±0.38比1.00±0.12,P<0.05),miR-4458表达量较癌旁组织降低(0.34±0.04比1.00±0.07,P<0.05);抑制si-CircDONSON或过表达miR-4458降低肝癌细胞活力、划痕愈合率和MMP-2、MMP-9蛋白水平(P<0.05)减少,细胞克隆形成数和侵袭细胞数(P<0.05);miR-4458过表达可降低WT-CircDONSON的荧光素酶活性(P<0.05);干扰miR-4458表达逆转了抑制CircDONSON表达对肝癌Huh-7细胞增殖、迁移、侵袭的作用(P<0.05)。结论:抑制CircDONSON表达可阻碍肝癌细胞增殖、克隆形成、迁移及侵袭,这可能与抑制CircDONSON导致细胞中miR-4458表达升高有关。
Objective:To explore the effect of CircDONSON on the proliferation,migration and invasion of liver cancer Huh-7 cells and its possible mechanism.Methods:The expression levels of CircDONSON and miR-4458 in liver cancer tissues were detected by qRT-PCR;liver cancer cells Huh-7 were divided into si-NC group,si-CircDONSON group,miR-NC group,miR-4458 group,si-CircDONSON+anti-miR-NC group,si-CircDONSON+anti-miR-4458 group;CCK-8 method,plate clone formation assay,scratch assay and Transwell assay were used to detect cell proliferation,clone formation,migration and invasion;dual-luciferase reporter assay to detect the targeting of miR-4458 and CircDONSON;Western blot to detect the protein expression of MMP-2 and MMP-9.Results:The expression of CircDONSON in liver cancer tissues was higher than that in adjacent tissue(4.82±0.38 vs 1.00±0.12,P<0.05),but the expression of miR-4458 was lower than that in adjacent tissue(0.34±0.04 vs 1.00±0.07,P<0.05).Inhibition of si-CircDONSON or overexpression of miR-4458 reduced liver cancer cell viability,scratch healing rate,MMP-2 and MMP-9 protein levels(P<0.05),and the number of cell clones and invasive cells(P<0.05);Overexpression of miR-4458 could reduce the luciferase activity of WT-CircDONSON(P<0.05);interference with miR-4458 expression reversed the effect of inhibiting CircDONSON expression on the proliferation,migration and invasion of liver cancer Huh-7 cells(P<0.05).Conclusion:Inhibition of CircDONSON expression could hinder the proliferation,clone formation,migration and invasion of liver cancer cells,which may be related to the increased expression of miR-4458 in cells caused by inhibition of CircDONSON.
作者
吕家誉
王宇
史云丰
赵琳
LYU Jia-yu;WANG Yu;SHI Yun-feng;ZHAO Lin(Department of General Surgery,Harbin Fifth Hospital,Harbin 150040,China;Department of Orthopedics,Harbin Fourth Hospital,Harbin 150040,China;Department of Emergency,Harbin Fifth Hospital,Harbin 150040,China)
出处
《中国现代普通外科进展》
CAS
2023年第5期354-357,362,共5页
Chinese Journal of Current Advances in General Surgery
