LncRNA SOX2-OT靶向miR-186-5p/ZEB1轴对黑色素瘤细胞增殖、迁移和侵袭的影响OA
Effects of LncRNA SOX2-OT on Proliferation,Migration,and Invasion of Melanoma Cells by Targeting the miR-186-5p/ZEB1 Axis
目的 探讨长链非编码RNA SOX2重叠转录本(LncRNA SOX2-OT)靶向微小RNA-186-5p(miR-186-5p)/锌指E盒结合同源框1(ZEB1)轴对黑色素瘤(MM)细胞的影响.方法 体外常规培养正常人皮肤黑色素细胞系PIG1以及人 MM 细胞系 A375、M14 和 A2058,检测各细胞 LncRNA SOX2-OT、miR-186-5p、ZEB1 mRNA 表达水平.将 A2058 细胞分为对照组、si-NC 组、si-SOX2-OT 组、si-SOX2-OT+inhibitor-NC 组、si-SOX2-OT+miR-186-5p inhibitor 组.检测 LncRNA SOX2-OT、miR-186-5p、ZEB1 mRNA表达水平(RT-qPCR法);细胞活力(CCK-8试剂盒法)、增殖活性(克隆形成实验)、迁移(划痕试验)、侵袭能力(Transwell小室法);ZEB1、E-cadherin、TGF-β1、α-SMA蛋白表达量(Western blot法).验证miR-186-5p与LncRNA SOX2-OT、ZEB1之间的靶向关系.结果 与对照组、si-NC组比较,si-SOX2-OT组LncRNA SOX2-OT、ZEB1 mRNA表达水平、OD450值、细胞克隆数、划痕愈合率、细胞侵袭数以及ZEB1、TGF-β1和α-SMA蛋白表达量降低,miR-186-5p表达水平以及E-cadherin蛋白表达量升高(P＜0.05);与si-SOX2-OT+inhibitor-NC组比较,si-SOX2-OT+miR-186-5p inhibitor组ZEB1 mRNA水平、OD450值、细胞克隆数、划痕愈合率、细胞侵袭数以及ZEB1、TGF-β1和α-SMA蛋白表达量升高,miR-186-5p表达水平及E-cadherin蛋白表达量降低(P＜0.05).转染miR-186-5p mimic后,LncRNA SOX2-OT-WT、ZEB1-WT的荧光素酶活性降低(P＜0.05).裸鼠移植瘤实验显示,敲低SOX2-OT可提高移植瘤miR-186-5p和E-cadherin蛋白表达,降低ZEB1蛋白表达、抑制移植瘤的生长;抑制miR-186-5p的表达可减轻敲低SOX2-OT对移植瘤生长的抑制作用(P＜0.05).结论 在MM细胞中LncRNA SOX2-OT异常高表达,敲低LncRNA SOX2-OT可靶向上调miR-186-5p的表达,抑制ZEB1的表达,进而抑制MM细胞的恶性生物学行为.
Objective To discuss effects of long non-coding small nucleolar RNA SOX2 overlapping transcript(LncRNA SOX2-OT)on melanoma(MM)cells by targeting microRNA-186-5p(miR-186-5p)/zinc finger E-box binding homeobox protein 1(ZEB1)axis.Methods Normal human skin melanocyte line PIG1 and human MM cell lines A375,M14,and A2058 were cul-tured in vitro,and the LncRNA SOX2-OT,miR-186-5p,and ZEB1 mRNA in each cell were detected.A2058 cells were separated into Control group,si-NC group,si-SOX2-OT group,si-SOX2-OT+inhibitor-NC group,and si-SOX2-OT+miR-186-5p inhibitor group.The LncRNA SOX2-OT,miR-186-5p,ZEB1 mRNA(RT-qPCR method),cell viability(CCK-8 assay kit),proliferation ac-tivity(clone formation assay),migration(scratch assay),invasion ability(Transwell chamber assay),ZEB1,E-cadherin,TGF-β1,and α-SMA proteins were detected by Western blot.Targeting relationships between miR-186-5p and LncRNA SOX2-OT,ZEB1 were validated.Results Compared with control group and si-NC group,si-SOX2-OT group showed a decrease in LncRNA SOX2-OT and ZEB1 mRNAs,OD450 value,cell clone number,scratch healing rate,cell invasion number,and ZEB1,TGF-β1,andα-SMA proteins,as well as an increase in miR-186-5p and E-cadherin protein(P＜0.05).Compared with si-SOX2-OT+inhibi-tor-NC group,si-SOX2-OT+miR-186-5p inhibitor group showed an increase in ZEB1 mRNA,OD450 value,cell clone number,scratch healing rate,cell invasion number,and ZEB1,TGF-β1,and α-SMA proteins,and a decrease in miR-186-5p and Ecadherin protein(P＜0.05).After transfection with miR-186-5p mimic,the luciferase activity of LncRNA SOX2-OT-WT and ZEB1-WT de-creased(P＜0.05).The nude mouse transplantation experiment showed that knocking down SOX2-OT could increase the miR-186-5p and E-cadherin proteins in transplanted tumors,reduce ZEB1 protein,and inhibit the growth of transplanted tumors.Inhibiting miR-186-5p could alleviate the inhibitory effect of knocking down SOX2-OT on the growth of transplanted tumors(P＜0.05).Conclusion LncRNA SOX2-OT is abnormally overexpressed in MM cells.Knocking down LncRNA SOX2-OT could target upregulation of miR-186-5p,inhibit ZEB1,and thus suppress the malignant biological behaviors of MM cells.
邱文娜;张龙军;冯金;刘波
邢台市人民医院妇科,整形外科,河北邢台 054000邢台市人民医院妇科,整形外科,河北邢台 054000邢台市人民医院妇科,整形外科,河北邢台 054000邢台市人民医院妇科,整形外科,河北邢台 054000
医药卫生
长链非编码RNA SOX2重叠转录本微小RNA-186-5p/锌指E盒结合同源框1轴黑色素瘤细胞恶性生物学行为
long non-coding RNA SOX2 overlapping transcriptmicroRNA-186-5p/zinc finger E-box binding homeobox protein 1melanoma cellsmalignant biological behaviors
《四川医学》 2026 (3)
272-278,7
河北省卫生健康委员会项目资助(编号:20251413)
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