lncRNA HCP5靶向miR-409-3p对皮肤鳞状细胞癌细胞增殖、迁移和侵袭的影响

lncRNA HCP5 targets miR-409-3p to regulate the proliferation,migration and invasion of cutaneous squamous cell carcinoma cells

目的:探讨lncRNA HCP5靶向mlR-409-3p对皮肤鳞状细胞癌(CSCC)细胞增殖、迁移和侵袭的影响。方法:采用qRT-PCR法检测CSCC细胞(SCC13、A431、HSC-5)中HCP5和miR-409-3p的表达情况,筛选用于实验的CSCC细胞。将抑制HCP5表达的质粒si-HCP5及其对照(si-NC)、过表达miR-409-3p的miR-409-3p模拟物及其对照(miR-NC)、抑制HCP5和miR-409-3p的si-HCP5+anti-miR-409-3p及其对照(si-HCP5+anti-miR-NC)分别转染SCC13细胞。采用CCK-8法和Transwell实验检测SCC13细胞增殖、迁移、侵袭能力,Western blot法检测细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶(MMP)2、MMP9蛋白表达水平。通过双荧光素酶报告基因实验验证HCP5和miR-409-3p的靶向关系。结果:SCC13细胞中HCP5表达最高,miR-409-3p表达最低(P<0.05)。抑制HCP5表达后SCC13细胞增殖能力降低,细胞迁移数和侵袭数减少,CycIinD1、MMP2、MMP9蛋白表达水平降低(P<0.05)。过表达miR-409-3p后SCC13细胞增殖能力降低,细胞迁移数和侵袭数减少,CycinD1、MMP2、MMP9蛋白表达水平降低(P<0.05)。双荧光素酶报告基因实验结果显示lncRNA HCP5和miR-409-3p存在靶向关系。抑制miR-409-3p表达可减弱HCP5低表达对SCC13细胞增殖以及迁移、侵袭能力的影响(P<0.05)。结论:抑制HCP5可通过靶向促进miR-409-3p表达来抑制CSCC细胞增殖、迁移和侵袭。

Aim:To explore the effects of lncRNA HCP5 targeting by miR-409-3p on the proliferation,migration and invasion of skin phosphor cell carcinoma(CSCC)cells.Methods:The expression of HCP5 and miR-409-3p in CSCC cells(SCC13、A431、HSC-5)was detected by qRT-PCR,and the CSCC cells used in the experiment were screened.The plasmid si-HCP5 inhibiting the expression of HCP5 and its control(si-NC),miR-409-3p mimics with overexpression of miR-409-3p and its control(miR-NC),si-HCP5+anti-miR-409-3p and its control(si-HCP5+anti-miR-NC)were transfected into SCC13 cells.CCK-8 assay and Transwell assay were used to detect the proliferation,migration and invasion ability of SCC13 cells,and Western blot assay was used to detect the expression levels of CyclinD1,MMP2 and MMP9 proteins.Dual luciferase reporter gene experiment was used to verify the targeting relationship between HCP5 and miR-409-3p.Results:The expression of HCP5 was highest,while miR-409-3p was lowest in SCC13 cells(P<0.05).After inhibiting the expression of HCP5,the proliferation of SCC13 cells was decreased,and the number of migration and invasion were significantly decreased,the expression of CyclinD1,MMP2,MMP9 was decreased(P<0.05).After overexpression of miR-409-3p,the proliferation of SCC13 cells was decreased,and the number of migration and invasion were significantly decreased,and the expression levels of CycinD1,MMP2 and MMP9 proteins were decreased(P<0.05).The results of dual luciferase reporter gene experiment showed that there was a targeting relationship between HCP5 and miR-409-3p.Inhibiting the expression of miR-409-3p could reduce the effects of low expression of HCP5 on the proliferation,migration and invasion of SCC13 cells(P<0.05).Conclusion:Inhibition of HCP5 can inhibit the proliferation,migration and invasion of CSCC cells by targeting and promoting the expression of miR-409-3p.