LncRNA DSCAM-AS1调节miR-431-5p/PRDX1信号轴对肝癌细胞恶性进展的影响  

LncRNA DSCAM-AS1 regulates the miR-431-5p/PRDX1 signaling axis to influence the malignant progression of hepatocellular carcinoma cells

作　　者：陈志飞[1] 孙伟涛[2] 颜廷启[2] 孙江江[2] 石彦科 于生才 黄伟 CHEN Zhi-fei;SUN Wei-tao;YAN Ting-qi;SUN Jiang-jiang;SHI Yan-ke;YU Sheng-cai;HUANG Wei(First Department of Emergency Surgery,Handan Central Hospital,Handan 056004,Hebei,China;不详)

机构地区：[1]邯郸市中心医院急诊外一科,河北邯郸056004 [2]邯郸市中心医院普外十科,河北邯郸056004 [3]江南大学附属医院普外科,江苏无锡214122

出　　处：《广东医学》2025年第2期167-175,共9页Guangdong Medical Journal

基　　金：河北省医学科学研究课题(20220513)。

摘　　要：目的探究长链非编码RNA(LncRNA)DSCAM-AS1调节微小RNA-431-5p(miR-431-5p)/过氧化物氧化还原蛋白1(PRDX1)信号轴对肝癌细胞增殖、凋亡、迁移、侵袭和上皮间质转化(EMT)的影响。方法实时荧光定量聚合酶链式反应(qRT-PCR)检测人正常肝细胞系L02以及人肝癌细胞系Hep G2中LncRNA DSCAM-AS1、miR-431-5p水平;将si-DSCAM-AS1+miR-431-5p inhibitor、si-DSCAM-AS1+inhibitor NC、si-DSCAM-AS1、si-NC分别转染至Hep G2细胞并记为si-DSCAM-AS1+miR-431-5p inhibitor组、si-DSCAM-AS1+inhibitor NC组、si-DSCAM-AS1组、si-NC组,未作处理的Hep G2细胞记为NC组。双荧光素酶报告基因实验验证LncRNA DSCAM-AS1与miR-431-5p、PRDX1的关系;qRT-PCR检测Hep G2细胞中LncRNA DSCAM-AS1、miR-431-5p表达;噻唑蓝(MTT)法检测Hep G2细胞增殖情况;流式细胞术检测Hep G2细胞凋亡率;Transwell检测Hep G2细胞侵袭、迁移数量;蛋白质印迹法(Western blot)检测Hep G2细胞Ki-67、神经型钙黏附蛋白(N-cadherin)、波形蛋白(Vimentin)、上皮钙黏附素(E-cadherin)、PRDX1蛋白表达。结果与L02细胞相比,Hep G2细胞LncRNA DSCAM-AS1表达水平、PRDX1蛋白表达显著上调(P<0.05),miR-431-5p表达水平显著下调(P<0.05)。与NC组、si-NC组相比,si-DSCAM-AS1组Hep G2细胞A570、LncRNA DSCAM-AS1表达、侵袭细胞数量、N-cadherin、迁移细胞数量、Vimentin蛋白表达、Ki-67显著下降(P<0.05),Hep G2细胞凋亡率、miR-431-5p表达水平、E-cadherin蛋白表达显著升高(P<0.05);而抑制miR-431-5p表达削弱了沉默LncRNA DSCAM-AS1抑制Hep G2细胞增殖、迁移、侵袭的作用;LncRNA DSCAM-AS1负向调控miR-431-5p/PRDX1轴。结论沉默LncRNA DSCAM-AS1可能通过上调miR-431-5p来下调PRDX1的表达,诱导细胞凋亡,并抑制肝癌细胞增殖和EMT,进而抑制肝癌细胞系Hep G2的恶性进展。Objective To explore the effects of long non-coding RNA(LncRNA)DSCAM-AS1 on hepatocellular carcinoma(HCC)cell proliferation,apoptosis,migration,invasion,and epithelial-mesenchymal transition(EMT)through the miR-431-5p/peroxiredoxin 1(PRDX1)signaling axis.Methods Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of LncRNA DSCAM-AS1 and miR-431-5p in the normal human hepatocyte cell line(L02)and the human hepatocellular carcinoma cell line(Hep G2).Hep G2 cells were transfected with si-DSCAM-AS1+miR-431-5p inhibitor,si-DSCAM-AS1+inhibitor NC,si-DSCAM-AS1,and si-NC were designated as the si-DSCAM-AS1+miR-431-5p inhibitor group,si-DSCAM-AS1+inhibitor NC group,si-DSCAM-AS1 group,and si-NC group,respectively.Untreated Hep G2 cells were considered the NC group.The interaction between LncRNA DSCAM-AS1,miR-431-5p,and PRDX1 was validated through dual-luciferase reporter assays.qRT-PCR was also used to assess LncRNA DSCAM-AS1 and miR-431-5p expression in Hep G2 cells.Cell proliferation was measured by MTT assay,apoptosis by flow cytometry,invasion and migration by Transwell assays,and protein expression(Ki-67,N-cadherin,Vimentin,E-cadherin,and PRDX1)by Western blot.Results Compared to L02 cells,Hep G2 cells exhibited significantly higher levels of LncRNA DSCAM-AS1 and PRDX1 expression(P<0.05),whereas miR-431-5p levels were markedly reduced(P<0.05).Compared to the NC and si-NC groups,the si-DSCAM-AS1 group demonstrated significantly reduced A570 absorbance,LncRNA DSCAM-AS1 expression,invasive cell numbers,N-cadherin,migrating cell numbers,Vimentin protein expression,and Ki-67(P<0.05).Additionally,the si-DSCAM-AS1 group showed significantly increased apoptosis rates,miR-431-5p expression,and E-cadherin protein levels(P<0.05).Notably,inhibiting miR-431-5p expression attenuated the suppressive effects of LncRNA DSCAM-AS1 silencing on Hep G2 cell proliferation,migration,and invasion.LncRNA DSCAM-AS1 was found to negatively regulate the miR-431-5p/PRDX1 axis.Conclusion Silencing LncRNA DSCA

关 键 词：LncRNA DSCAM-AS1 miR-431-5p/PRDX1轴 肝癌 增殖 凋亡 上皮间质转化 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤] R730.23[医药卫生—临床医学]