Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinoma

BackgroundAbnormal activation of the hepatocyte growth factor (HGF) and c-mesenchymal–epithelial transition factor (c-Met) signaling pathway is associated with tumor occurrence and development. Serum HGF concentrations are significantly higher in patients with advanced and poorly differentiated lary...

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Main Authors: Yixuan Huo, Wei Zhang, Fan Yang, Wenhua Shao, Guozheng Cong, Shoukai Zhang
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Oncology
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Online Access:https://www.frontiersin.org/articles/10.3389/fonc.2025.1553626/full
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author Yixuan Huo
Wei Zhang
Fan Yang
Wenhua Shao
Guozheng Cong
Shoukai Zhang
Shoukai Zhang
author_facet Yixuan Huo
Wei Zhang
Fan Yang
Wenhua Shao
Guozheng Cong
Shoukai Zhang
Shoukai Zhang
author_sort Yixuan Huo
collection DOAJ
description BackgroundAbnormal activation of the hepatocyte growth factor (HGF) and c-mesenchymal–epithelial transition factor (c-Met) signaling pathway is associated with tumor occurrence and development. Serum HGF concentrations are significantly higher in patients with advanced and poorly differentiated laryngeal squamous cell carcinoma than those with early and highly differentiated disease. NK4, a splice variant of HGF, can competitively bind to c-Met and acts as a specific antagonist of HGF. Although preliminary research has been conducted on the tumor-suppressing function of the NK4 gene, its specific mechanism of action in laryngeal cancer remains unclear.MethodsStable laryngeal squamous cell carcinoma cell lines expressing NK4 were developed using a lentiviral packaging method. The experimental group was labeled with PLV-NK4-TU212, whereas the control group was labeled with PLV-NC-TU212. Western blotting verified a stable expression. The functions of the NK4 molecule were assessed using MTT, EMT, and apoptosis assays, and cell lines were subjected to transcriptome sequencing.ResultsProtein expression analysis showed that NK4 was stably expressed. Compared with the wild-type and negative control groups, overexpression of the NK4 gene inhibited the migration and proliferation of laryngeal squamous cell carcinoma cells and induced cell apoptosis. Transcriptome sequencing revealed that the expression levels of 320 genes differed significantly, with 189 upregulated and 131 downregulated genes.ConclusionIn this study, a TU212 laryngeal squamous cell carcinoma cell line overexpressing NK4 was constructed using a lentiviral packaging system. Functional experiments showed that PLV-NK4-TU212 cells exhibited a significantly reduced migration rate, decreased proliferative ability, and increased apoptosis rate. The results of this study provide an experimental basis for NK4 as a potential therapeutic target for laryngeal squamous cell carcinoma highlighting its translational medical value.
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spelling doaj-art-5f5e1553e3e8472f945845d3cc5ca0822025-08-20T03:15:57ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2025-08-011510.3389/fonc.2025.15536261553626Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinomaYixuan Huo0Wei Zhang1Fan Yang2Wenhua Shao3Guozheng Cong4Shoukai Zhang5Shoukai Zhang6First Clinical Medical College, Ningxia Medical University, Yinchuan, ChinaLanzhou University, Lanzhou, Gansu, ChinaLanzhou University, Lanzhou, Gansu, ChinaLanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaLanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaFirst Clinical Medical College, Ningxia Medical University, Yinchuan, ChinaGansu Provincial Hospital, Lanzhou, Gansu, ChinaBackgroundAbnormal activation of the hepatocyte growth factor (HGF) and c-mesenchymal–epithelial transition factor (c-Met) signaling pathway is associated with tumor occurrence and development. Serum HGF concentrations are significantly higher in patients with advanced and poorly differentiated laryngeal squamous cell carcinoma than those with early and highly differentiated disease. NK4, a splice variant of HGF, can competitively bind to c-Met and acts as a specific antagonist of HGF. Although preliminary research has been conducted on the tumor-suppressing function of the NK4 gene, its specific mechanism of action in laryngeal cancer remains unclear.MethodsStable laryngeal squamous cell carcinoma cell lines expressing NK4 were developed using a lentiviral packaging method. The experimental group was labeled with PLV-NK4-TU212, whereas the control group was labeled with PLV-NC-TU212. Western blotting verified a stable expression. The functions of the NK4 molecule were assessed using MTT, EMT, and apoptosis assays, and cell lines were subjected to transcriptome sequencing.ResultsProtein expression analysis showed that NK4 was stably expressed. Compared with the wild-type and negative control groups, overexpression of the NK4 gene inhibited the migration and proliferation of laryngeal squamous cell carcinoma cells and induced cell apoptosis. Transcriptome sequencing revealed that the expression levels of 320 genes differed significantly, with 189 upregulated and 131 downregulated genes.ConclusionIn this study, a TU212 laryngeal squamous cell carcinoma cell line overexpressing NK4 was constructed using a lentiviral packaging system. Functional experiments showed that PLV-NK4-TU212 cells exhibited a significantly reduced migration rate, decreased proliferative ability, and increased apoptosis rate. The results of this study provide an experimental basis for NK4 as a potential therapeutic target for laryngeal squamous cell carcinoma highlighting its translational medical value.https://www.frontiersin.org/articles/10.3389/fonc.2025.1553626/fullapoptosisgene expressionlaryngeal squamous cell carcinomaNK4RNA-seqmigration
spellingShingle Yixuan Huo
Wei Zhang
Fan Yang
Wenhua Shao
Guozheng Cong
Shoukai Zhang
Shoukai Zhang
Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinoma
Frontiers in Oncology
apoptosis
gene expression
laryngeal squamous cell carcinoma
NK4
RNA-seq
migration
title Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinoma
title_full Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinoma
title_fullStr Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinoma
title_full_unstemmed Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinoma
title_short Overexpression of NK4 gene in TU212 affects migratory activity in laryngeal squamous cell carcinoma
title_sort overexpression of nk4 gene in tu212 affects migratory activity in laryngeal squamous cell carcinoma
topic apoptosis
gene expression
laryngeal squamous cell carcinoma
NK4
RNA-seq
migration
url https://www.frontiersin.org/articles/10.3389/fonc.2025.1553626/full
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