中国耳鼻咽喉颅底外科杂志2024,Vol.30Issue(3) :9-15.DOI:10.11798/j.issn.1007-1520.202423293

长链非编码RNA核富集转录体1促进喉鳞状细胞癌细胞迁移和侵袭的机制研究

Mechanism research of long non-coding RNA nuclear enriched transcriptome 1 on the migration and invasion of laryngeal squamous cell carcinoma cells

梁冀望 高小卓 杨骁 孙铭 赵月皎
中国耳鼻咽喉颅底外科杂志2024,Vol.30Issue(3) :9-15.DOI:10.11798/j.issn.1007-1520.202423293

长链非编码RNA核富集转录体1促进喉鳞状细胞癌细胞迁移和侵袭的机制研究

Mechanism research of long non-coding RNA nuclear enriched transcriptome 1 on the migration and invasion of laryngeal squamous cell carcinoma cells

梁冀望 1高小卓 2杨骁 1孙铭 1赵月皎1
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作者信息

  • 1. 辽宁省肿瘤医院中国医科大学肿瘤医院头颈外科,辽宁沈阳 110042
  • 2. 辽宁省肿瘤医院中国医科大学肿瘤医院病理科,辽宁沈阳 110042
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摘要

目的 探究长链非编码RNA(lncRNA)核富集转录体1(NEAT1)NEAT1对喉鳞状细胞癌(LSCC)细胞迁移和侵袭的影响,并进一步分析miR-429/锌指E-盒结合同源异形盒1(ZEB1)轴在其中发挥的作用.方法 慢病毒转染建立稳定敲减lncRNA NEAT1的LSCC细胞,通过RT-qPCR检测细胞lncRNA NEAT1表达以验证转染效率,通过Transwell实验检测细胞迁移和侵袭能力,通过Western blot检测细胞上皮-间充质转化(EMT)相关蛋白N-cadherin、Vimentin、Slug和Snail表达.通过生物信息学分析miR-429与lncRNA NEAT1和ZEB1 mRNA间的潜在结合位点,并通过双荧光素酶报告基因实验验证miR-429与lncRNA NEAT1以及miR-429与ZEB1 mRNA结合.将miR-429抑制剂转染敲减lncRNA NEAT1的LSCC细胞,通过Western blot检测细胞ZEB1表达.进一步检查敲减ZEB1对抑制miR-429的敲减lncRNA NEAT1的LSCC细胞迁移、侵袭和EMT的影响.结果 敲减lncRNA NEAT1降低LSCC细胞lncRNA NEAT1表达,抑制细胞迁移和侵袭并下调细胞N-cadherin、Vimentin、Slug和Snail表达.miR-429 与 lncRNA NEAT1 和 ZEB1 mRNA 间存在潜在结合位点,且 miR-429 与 lncRNA NEAT1 以及 miR-429 与ZEB1 mRNA结合.抑制miR-429逆转了敲减lncRNA NEAT1对LSCC细胞ZEB1表达的抑制作用.此外,敲减ZEB1逆转了抑制miR-429对敲减lncRNA NEAT1的LSCC细胞迁移和侵袭的促进作用及EMT相关蛋白N-cadher-in、Vimentin、Slug和Snail表达的上调作用.结论 lncRNA NEAT1促进LSCC细胞迁移和侵袭,其机制可能与海绵化miR-429上调ZEB1表达促进LSCC细胞EMT有关.

Abstract

Objective To explore the effect of long non-coding RNA nuclear enriched transcriptome 1(lncRNA NEAT1)on the migration and invasion of laryngeal squamous cell carcinoma(LSCC)cells,and further explore the role of miR-429/zinc finger E-box binding homeobox 1(ZEB1)axis in this process.Methods Lentivirus transfection established LSCC cells with stable knockdown lncRNA NEAT1,and lncRNA NEAT1 expression was detected by real-time quantitative polymerase chain reaction(RT-qPCR)to verify transfection efficiency.Cell migration and invasion ability were detected by transwell assay.The expressions of epithelial-mesenchymal transition(EMT)related proteins N-cadherin,Vimentin,Slug and Snail were detected by Western blot.The potential binding sites between miR-429 and lncRNA NEAT1 and ZEB1 mRNA were analyzed by bioinformatics,and the binding of miR-429 to lncRNA NEAT1 and miR-429 to ZEB1 mRNA were verified by dual luciferase reporter gene assay.LSCC cells with knockdown lncRNA NEAT1 were transfected with miR-429 inhibitors,and ZEB1 expression was detected by Western blot.The effect of knockdown ZEB1 on LSCC cell migration,invasion and EMT of miR-429 knockdown lncRNA NEAT1 was further examined.Results Knocking down lncRNA NEAT1 reduced lncRNA NEAT1 expression in LSCC cells,inhibited cell migration and invasion,and down-regulated the expression of N-cadherin,Vimentin,Slug,and Snail.There were potential binding sites between miR-429 and lncRNA NEAT1 and ZEB1 mRNA,and miR-429 bound to lncRNA NEAT1 and miR-429 to ZEB1 mRNA.Inhibition of miR-429 reversed the inhibitory effect of knocking down lncRNA NEAT1 on ZEB1 expression in LSCC cells.In addition,knockdown of ZEB1 reversed the inhibitory effect of miR-429 on the migration and invasion of LSCC cells with knockdown lncRNA NEAT1,as well as the upregulation of EMT related proteins N-cadherin,Vimentin,Slug,and Snail expression.Conclusion LncRNA NEAT1 promotes migration and invasion of LSCC cells,and the mechanism may be related to the upregulation of ZEB1 expression by sponging miR-429 and promoting EMT in LSCC cells.

关键词

喉鳞状细胞癌/LncRNA/NEAT1/miR-429/ZEB1轴/细胞迁移和侵袭/上皮-间充质转化

Key words

Laryngeal squamous cell carcinoma/LncRNA NEAT1/MiR-429/ZEB1 axis/Cell migration and invasion/Epithelial-mesenchymal transformation

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基金项目

辽宁省自然科学基金(2023-MS-059)

出版年

2024
中国耳鼻咽喉颅底外科杂志
中南大学

中国耳鼻咽喉颅底外科杂志

CSTPCDCSCD
影响因子:0.643
ISSN:1007-1520
参考文献量2
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