LncRNA LUCAT1介导的miR-199b-5p/MAPKAPK3轴在甲状腺乳头状癌发展中的调控作用及机制

The regulatory role and mechanism of LncRNA LUCAT1 mediated miR-199b-5p/MAPKAPK3 axis in the development of papillary thyroid cancer

目的探究lncRNA LUCAT1对甲状腺乳头状癌TPC-1细胞增殖、迁移、侵袭、凋亡和EMT的影响,并进一步探究miR-199b-5p/MAPKAPK3轴在其中发挥的作用。方法RT-qPCR检测甲状腺乳头状癌组织及癌旁组织中LUCAT1、miR-199b-5p和MAPKAPK3表达;生物信息学预测miR-199b-5p与LUCAT1或MAPKAPK3的潜在结合位点,双荧光素酶报告基因实验验证miR-199b-5p与LUCAT1或MAPKAPK3结合。将TPC-1细胞分为sh-NC组、sh-LUCAT1组、sh-LUCAT1+inhibitor NC组、sh-LUCAT1+miR-199b-5p inhibitor组和sh-LUCAT1+miR-199b-5p inhibitor+sh-MAPKAPK3组;Western blot检测各组细胞MAPKAPK3蛋白表达水平;CCK-8实验检测各组细胞增殖活性;流式细胞术检测各组细胞凋亡率;Transwell实验检测各组细胞迁移和侵袭能力;Western blot方法检测各组细胞EMT相关蛋白表达水平。结果甲状腺乳头状癌组织中LUCAT1和MAPKAPK3高表达,且miR-199b-5p低表达;miR-199b-5p与LUCAT1和MAPKAPK3结合;敲减LUCAT1降低TPC-1细胞MAPKAPK3表达,抑制TPC-1细胞增殖、迁移、侵袭和EMT,并促进细胞凋亡;抑制miR-199b-5p逆转了敲减LUCAT1对TPC-1细胞增殖、迁移、侵袭和EMT的抑制作用及细胞凋亡的促进作用;敲减MAPKAPK3逆转了抑制miR-199b-5p对敲减LUCAT1的TPC-1细胞增殖、迁移、侵袭和EMT的促进作用及细胞凋亡的抑制作用。结论lncRNALUCAT1可能通过竞争结合miR-199b-5p上调MAPKAPK3表达促进甲状腺乳头状癌细胞增殖、迁移、侵袭和EMT并抑制细胞凋亡。

Objective To explore the effects of lncRNA LUCAT1 on the proliferation,migration,invasion,apoptosis,and EMT of papillary thyroid cancer TPC-1 cells,and further investigate the role of miR-199b-5p/MAPKAPK3 axis in this process.Methods The expressions of LUCAT1,miR-199b-5p,and MAPKAPK3 in papillary thyroid carcinoma and adjacentnormal tissues were detected by RT-qPCR.Bioinformatics predicted the potential binding sites of miR-199b-5p with LUCAT1 or MAPKAPK3,and verified the binding of miR-199b-5p with LUCAT1 or MAPKAPK3 viadouble luciferase reporter gene experiment.TPC-1 cells were divided into sh-NC group,sh-LUCAT1 group,sh-LUCAT1+inhibitor NC group,sh-LUCAT1+miR-199b-5p inhibitor group,and sh-LUCAT1+miR-199b-5p inhibitor+sh-MAPKAPK3 group.The expression of MAPKAPK3 of cells in each group were detected by Western blot.Cell proliferation activity of cells in each group were detected by CCK-8.Apoptosis rate of cells in each group were detected by flow cytometry.The ability of migration and invasion of cells in each group were detected by transwell assay.The expressions of EMT related proteins of cells in each group were detected by Western blot.Results LUCAT1 and MAPKAPK3 were high expression and miR-199b-5p was low expression in papillary thyroid carcinoma tissues.MiR-199b-5p binds to LUCAT1 and MAPKAPK3.Knocking down LUCAT1 reduced the expression of MAPKAPK3,inhibited cell proliferation,migration,invasion,and EMT,and promoted cell apoptosis of TPC-1 cells.Inhibiting miR-199b-5p reversed the inhibitory effect of knockdown LUCAT1 on cell proliferation,migration,invasion,EMT,and the promoting effect of cell apoptosis of TPC-1 cells.Knocking down MAPKAPK3 reversed the inhibitory effect of miR-199b-5p on cell proliferation,migration,invasion,EMT,and apoptosis of TPC-1 cells with knockdown LUCAT1.Conclusion lncRNA LUCAT1 may promote proliferation,migration,invasion,EMT,and inhibit cell apoptosis by competitively binding to miR-199b-5p and upregulating MAPKAPK3 expression.