摘要
目的 探讨长链非编码RNA(lncRNA)MIR4435-2HG(MIR4435-2HG)对胆管癌细胞增殖、迁移、侵袭、凋亡的影响及其对微小RNA-376a-3p(miR-376a-3p)的调控作用。方法 qRT-PCR法检测人肝内胆管上皮细胞HIBEpic与人胆管癌细胞RBE中MIR4435-2HG、miR-376a-3p的表达。将si-NC、si-MIR4435-2HG、miR-NC、miR-376a-3p mimics、si-MIR4435-2HG联合anti-miR-NC、si-MIR4435-2HG联合anti-miR-376a-3p分别转染至RBE细胞,作为si-NC组、si-MIR4435-2HG组、miR-NC组、miR-376a-3p组、si-MIR4435-2HG+anti-miR-NC组、si-MIR4435-2HG+anti-miR-376a-3p组;采用MTT法、Transwell小室法及流式细胞仪分别检测细胞增殖、迁移、侵袭及凋亡情况;双荧光素酶报告基因实验验证MIR4435-2HG与miR-376a-3p的靶向关系。Western blot检测相关蛋白表达。结果 RBE细胞中MIR4435-2HG表达量升高(P<0.05),miR-376a-3p表达量降低(P<0.05)。与si-NC组比较,si-MIR4435-2HG组MIR4435-2HG表达、细胞活力及CyclinD1、MMP-2、MMP-9蛋白水平降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),细胞凋亡率升高(P<0.05);与miR-NC组比较,miR-376a-3p组细胞活力及CyclinD1、MMP-2、MMP-9蛋白水平降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),miR-376a-3p表达、细胞凋亡率升高(P<0.05)。MIR4435-2HG可靶向调控miR-376a-3p;与si-MIR4435-2HG+anti-miR-NC组比较,si-MIR4435-2HG+anti-miR-376a-3p组细胞活力及CyclinD1、MMP-2、MMP-9蛋白水平升高(P<0.05),迁移及侵袭细胞数增多(P<0.05),miR-376a-3p表达、细胞凋亡率降低(P<0.05)。结论 敲低MIR4435-2HG可通过靶向调控miR-376a-3p进而抑制RBE细胞增殖、迁移、侵袭,并诱导其凋亡。
Objective To explore the effects of long non-coding RNA(lncRNA)MIR4435-2HG(MIR4435-2HG)on the proliferation,migration,invasion and apoptosis of cholangiocarcinoma cells and its regulatory effect on microRNA-376a-3p(miR-376a-3p).Methods qRT-PCR method was used to detect the expression of MIR4435-2HG and miR-376a-3p in human intrahepatic bile duct epithelial cells HIBEpic and human cholangiocarcinoma cells RBE.si-NC,si-MIR4435-2HG,miR-NC,miR-376a-3p mimics,si-MIR4435-2HG and anti-miR-NC,and si-MIR4435-2HG and anti-miR-376a-3p were transfected into RBE cells,respectively,as the si-NC group,the si-MIR4435-2HG group,the miR-NC group,the miR-376a-3p group,the si-MIR4435-2HG+anti-miR-NC group,the si-MIR4435-2HG+anti-miR-376a-3p group.MTT method,Transwell chamber method and flow cytometry were used to detect cell proliferation,migration,invasion and apoptosis;dual luciferase reporter gene assay was used to verify the targeting relationship between MIR4435-2HG and miR-376a-3p.Western blot was used to detect the expression of related proteins.Results The expression of MIR4435-2HG was increased in RBE cells,while the expression of miR-376a-3p was decreased(P<0.05).Compared with the si-NC group,the MIR4435-2HG expression,cell viability,and protein levels of CyclinD1,MMP-2,MMP-9 in the si-MIR4435-2HG group were reduced(P<0.05),the numbers of migrating and invading cells were reduced(P<0.05),while the MIR4435-2HG expression and apoptosis rate were increased(P<0.05).Compared with the miR-NC group,the cell viability and protein levels of CyclinD1,MMP-2,MMP-9 in the miR-376a-3p group were decreased(P<0.05),the numbers of migrating and invading cells were decreased(P<0.05),while the MIR4435-2HG expression and apoptosis rate were increased(P<0.05).MIR4435-2HG was of targeted regulation on miR-376a-3p.Compared with the si-MIR4435-2HG+anti-miR-NC group,the cell viability and protein levels of CyclinD1,MMP-2,MMP-9 in the si-MIR4435-2HG+anti-miR-376a-3p group were increased(P<0.05),the numbers of migrating and invading cells were incre
作者
刘文东
张嘉麟
余紫丹
LIU Wen-dong;ZHANG Jia-lin;YU Zi-dan(Department of Oncology,Huizhou Third People's Hospital,Huizhou Guangdong516001,China;Rehabilitation Center,Huizhou Third People's Hospital,Huizhou Guangdong 516001,China)
出处
《局解手术学杂志》
2024年第1期30-35,共6页
Journal of Regional Anatomy and Operative Surgery
基金
惠州市医疗卫生类科技计划项目(20190401)。
