lncRNA FAM83H-AS1通过抑制miR-383-3p表达调控食管癌细胞增殖、迁移、侵袭的分子机制研究

Molecular mechanisms of lncRNA FAM83H-AS1 regulating proliferation,migration and invasion of esophageal cancer cells by inhibiting expression of miR-383-3p

目的:探讨lncRNA FAM83H-AS1对食管癌细胞增殖、迁移、侵袭的影响及分子机制。方法:实验设置si-NC组、si-FAM83H-AS1组、pcDNA组、pcDNA-FAM83H-AS1组、miR-NC组、miR-383-3p组、si-FAM83H-AS1+anti-miR-NC组、siFAM83H-AS1+anti-miR-383-3p组。实时荧光定量PCR(RT-qPCR)检测FAM83H-AS1和miR-383-3p的表达水平;四甲基偶氮唑盐比色法(MTT)检测细胞增殖活性;蛋白质印迹(Western blot)法检测CyclinD1、p21、MMP-2、MMP-9蛋白表达;Transwell检测细胞迁移和侵袭;荧光素酶报告实验检测FAM83H-AS1对miR-383-3p的靶向调控。结果:与癌旁组织相比,食管癌组织中FAM83H-AS1高表达,miR-383-3p低表达(P<0.05)。抑制FAM83H-AS1表达或过表达miR-383-3p,细胞活性显著降低,细胞迁移、侵袭数显著降低,CyclinD1、MMP-2、MMP-9表达水平显著降低,p21表达水平显著升高(P<0.05)。FAM83H-AS1靶向调控miR-383-3p,干扰miR-383-3p表达逆转了抑制lncRNA FAM83H-AS1表达对食管癌EC9706细胞增殖、迁移和侵袭的抑制作用。结论:抑制lncRNA FAM83H-AS1表达可能通过上调miR-383-3p表达抑制食管癌细胞增殖、迁移和侵袭。

Objective:To investigate the effect of lncRNA FAM83H-AS1 on the proliferation,migration and invasion of esophageal cancer cells and its molecular mechanism.Methods:Experiment set si-NC group,si-FAM83H-AS1 group,pcDNA group,pcDNA-FAM83H-AS1 group,miR-NC group,miR-383-3p group,si-FAM83H-AS1+anti-miR-NC group,si-FAM83H-AS1+anti-miR-383-3p group.Real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expressions of FAM83H-AS1 and miR-383-3p;tetramethylazozolium colorimetry(MTT)was used to detect cell proliferation activity;Western blot was used to detect CyclinD1,p21,MMP-2,MMP-9 protein expression;Transwell was used to detect cell migration and invasion;luciferase reporter assay to detect the targeted regulation between miR-383-3p and FAM83H-AS1.Results:Compared with adjacent tissues,FAM83H-AS1 was highly expressed and miR-383-3p lowly expressed in esophageal cancer tissues(P<0.05).Inhibition of FAM83H-AS1 expression or overexpression of miR-383-3p,cell viability was significantly reduced,the number of cell migration and invasion was significantly reduced,the expressions of CyclinD1,MMP-2,MMP-9 were significantly reduced,and the expression of p21 was significantly increased(P<0.05).FAM83H-AS1 targeted regulation of miR-383-3p,and interference with miR-383-3p expression reversed the inhibition of lncRNA FAM83H-AS1 expression on the proliferation,migration and invasion of esophageal cancer EC9706 cells.Conclusion:Inhibition of lncRNA FAM83H-AS1 expression may inhibit the proliferation,migration and invasion of esophageal cancer cells by up-regulating the expression of miR-383-3p.