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

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。

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

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-380-3p in human immortalized epidermal cells (HaCaT) and CSCC cells (SCC13,A431 and HSC-5).LINC00460 small interfering RNA (si-LINC00460),miR-380-3p mimics (miR-380-3p mimics),si-LINC00460+miR-380-3p inhibitors (anti-miR-380-3p) 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-380-3p.Results:Compared with HaCaT,the expression of LINC00460 in the CSCC cells was significantly increased,while the expression of miR-380-3p 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 proteins were significantly reduced (P＜0.05),apoptosis rate and E-Cadherin protein expression were significantly increased (P＜0.05).After up-regulation of miR-380-3p expression,the proliferation,migration and invasion of A431 cells,the expression of CyclinD1,MMP2 and MMP9 were significantly reduced (P＜0.05),apoptosis rate was significantly increased (P＜0.05).Compared with the down-regulation of LINC00460,the proliferation,migration and invasion of A431 cells,the expression of CyclinD1,MMP2,MMP9,N-Cadherin and Vimentin were significantly increased after down-regulation of LINC00460 and miR-380-3p (P＜0.05),while apoptosis rate and E-Cadherin protein expression were significantly reduced (P＜0.05).miR-380-3p was negatively regulated by LINC00460.Conclusion:The expression of LINC00460 is increased in CSCC cells,and down-regulation of LINC00460 could reduce the proliferation,migration and invasion ability and induce cell apoptosis of CSCC cells.The mechanism is related to the targeted regulation of miR-380-3p expression.