LINC00460靶向miR-380-3p对皮肤鳞状细胞癌细胞增殖、迁移、侵袭和凋亡的影响  

Effects of LINC00460 on the proliferation,migration,invasion and apoptosis of cutaneous squamous cells carcinoma by targeting miR-380-3p

作　　者：张翠[1] 王珺[1] 林伟[1] 孙田[1] ZHANG Cui;WANG Jun;LIN Wei;SUN Tian(Department of Dermatology,the Seventh People's Hospital of Shenyang,Liaoning Shenyang 110000,China)

机构地区：[1]沈阳市第七人民医院皮肤科,辽宁沈阳110000

出　　处：《现代肿瘤医学》2021年第24期4275-4281,共7页Journal of Modern Oncology

基　　金：辽宁省沈阳市卫计局科研计划项目(编号:2016688)。

摘　　要：目的:探讨LINC00460对皮肤鳞状细胞癌(CSCC)细胞增殖、迁移、侵袭和凋亡的影响及可能机制。方法:实时荧光定量PCR(RT-qPCR)分析人类永生化表皮细胞(HaCaT)和CSCC细胞(SCC13、A431和HSC-5)中LINC00460和miR-380-3p的表达水平。将LINC00460小干扰RNA(si-LINC00460)、miR-380-3p模拟物(miR-380-3p mimics)、si-LINC00460+miR-380-3p抑制物(anti-miR-380-3p)分别转染A431细胞,细胞计数试剂盒(CCK-8)检测细胞活力,流式细胞术分析细胞凋亡,Transwell实验分析细胞迁移和侵袭能力,蛋白质印记(Western blot)检测细胞周期素D1(CyclinD1)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)、上皮细胞钙黏蛋白(E-Cadherin)、神经钙黏蛋白(N-Cadherin)、波形蛋白(Vimentin)的表达。双荧光素酶报告基因实验、RT-qPCR确定LINC00460对miR-380-3p的靶向调控作用。结果:与HaCaT比较,CSCC细胞中LINC00460表达显著增加,miR-380-3p表达显著降低(P<0.05)。下调LINC00460表达后A431细胞增殖活力、迁移和侵袭细胞数以及CyclinD1、MMP2、MMP9、N-Cadherin和Vimentin蛋白表达显著降低(P<0.05),凋亡率、E-Cadherin蛋白表达显著升高(P<0.05)。上调miR-380-3p表达后A431细胞增殖活力、迁移和侵袭细胞数以及CyclinD1、MMP2和MMP9表达显著降低(P<0.05),凋亡率显著升高(P<0.05)。与下调LINC00460比较,同时下调LINC00460和miR-380-3p后A431细胞增殖活力、迁移和侵袭细胞数以及CyclinD1、MMP2、MMP9、N-Cadherin和Vimentin蛋白表达显著升高(P<0.05),凋亡率、E-Cadherin蛋白表达显著降低(P<0.05)。LINC00460靶向负调控miR-380-3p表达。结论:CSCC细胞中LINC00460表达增加,下调LINC00460能够降低CSCC细胞的增殖、迁移和侵袭能力,诱导细胞凋亡,其机制与靶向调控miR-380-3p表达有关。Objective:To explore the effect of LINC00460 on the proliferation,migration,invasion and apoptosis of cutaneous squamous cell carcinoma(CSCC)and its possible mechanism.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)was used to analyze the expression levels of LINC00460 and miR-380-3 p in human immortalized epidermal cells(HaCaT)and CSCC cells(SCC13,A431 and HSC-5).LINC00460 small interfering RNA(si-LINC00460),miR-380-3 p mimics(miR-380-3 p mimics),si-LINC00460+miR-380-3 p inhibitors(anti-miR-380-3 p)was transfected into A431 cells,respectively.Cell Counting Kit(CCK-8)detected cell viability,and Flow cytometry analyzed cell apoptosis.Transwell assay was analyzed cell migration and invasion ability.Western blot detected the expression of CyclinD1,matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9),E-Cadherin,N-Cadherin and Vimentin proteins.The dual luciferase report gene experiment and RT-qPCR confirmed the targeted regulation effect of LINC00460 on miR-380-3 p.Results:Compared with HaCaT,the expression of LINC00460 in the CSCC cells was significantly increased,while the expression of miR-380-3 p was significantly decreased(P<0.05).After down-regulating the expression of LINC00460,the proliferation,migration and invasion of A431 cells,the expression of CyclinD1,MMP2,MMP9,N-Cadherin and Vimentin proteinswere significantly reduced(P<0.05),apoptosis rate and E-Cadherin protein expression were significantly in-creased(P<0.05).After up-regulation of miR-380-3 p expression,the proliferation,migration and invasion ofA431 cells,the expression of CyclinD1,MMP2 and MMP9 were significantly reduced(P<0.05),apoptosis rate wassignificantly increased(P<0.05).Compared with the down-regulation of LINC00460,the proliferation,migrationand invasion of A431 cells,the expression of CyclinD1,MMP2,MMP9,N-Cadherin and Vimentin were significantlyincreased after down-regulation of LINC00460 and miR-380-3 p(P<0.05),while apoptosis rate and E-Cad-herin protein expression were significantly reduced(P<0.05).miR-380-3 p wa

关 键 词：LINC00460 皮肤鳞状细胞癌 增殖 迁移和侵袭 凋亡 miR-380-3p 

分 类 号：R739.5[医药卫生—肿瘤]