靶向沉默CXCL5对喉鳞状细胞癌AMC-HN-8细胞生物学功能的影响及其调控分析  

Effect of targeted CXCL5 silencing on biological function of laryngeal squamous cell carcinoma cell line AMC-HN-8 and its regulation analysis

作　　者：谢晓丹 韩瑞 卜倩倩 李梦璐 张亚楠 黄赛亚 皇甫辉 XIE Xiaodan;HAN Rui;BU Qianqian;LI Menglu;ZHANG Yanan;HUANG Saiya;HUANG Fuhui(The First Clinical Medical College of Shanxi Medical University,Taiyuan 030000,Shanxi Province,China)

机构地区：[1]山西医科大学第一临床医学院,山西太原030000

出　　处：《中国生物制品学杂志》2024年第1期17-25,共9页Chinese Journal of Biologicals

基　　金：山西省卫生计生委科研课题(2018044)。

摘　　要：目的 探讨靶向沉默CXCL5对喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)AMC-HN-8细胞相关生物学功能的影响,并通过TCGA数据库进行相关的调控分析。方法 在TCGA数据库中获取LSCC相关RNA-seq数据,分析CXCL5基因在LSCC及其癌旁组织中的表达差异;选取2019年1月—2020年12月山西医科大学第一医院60例LSCC及其癌旁组织样本,采用免疫组织化学染色法检测LSCC组织中CXCL5蛋白的表达;培养人LSCC细胞系AMCHN-8,qPCR法检测AMC-HN-8细胞中CXCL5 mRNA转录水平;筛选敲降效率较高的2组siRNA,CCK8法检测细胞增殖情况,Transwell试验检测细胞侵袭、迁移情况,流式细胞术检测细胞周期和凋亡情况;通过ssGSEA分析CXCL5与LSCC肿瘤免疫浸润水平的相关性,构建CXCL5共表达基因网络并对其进行GO和KEGG富集分析。结果 与癌旁组织及对照组细胞相比,CXCL5在LSCC组织及细胞中表达均升高,与TCGA数据库分析一致;在AMC-HN-8细胞中抑制CXCL5表达,具有抑制肿瘤细胞增殖、侵袭、迁移的能力,通过G1期抑制细胞周期,促进其凋亡;在DC、中性粒细胞(neutrophils)、NK和TH17免疫细胞评分存在差异。结论 CXCL5基因在LSCC组织中高表达,可能是LSCC靶向治疗的靶点之一。Objective To investigate the effects of targeted silencing of CXCL5 on the related biological functions of laryngeal squamous cell carcinoma(LSCC)cell line AMC-HN-8,and analyze the regulation through TCGA database.Methods RNA-seq data related to LSCC were obtained from TCGA database,and the expression differences of CXCL5 gene in LSCC and the adjacent tissues were analyzed.Total 60 samples of LSCC and the adjacent tissues from January 2019 to December 2020 were selected from the First Hospital of Shanxi Medical University,and the expression of CXCL5 protein in LSCC tissues was detected by immunohistochemical staining.Human LSCC cell line AMC-HN-8 was cultured and the mRNA transcription level of CXCL5 in AMC-HN-8 cells was detected by qPCR.Two groups of SiRNA with high knock-down efficiency were screened,CCK8 assay was used to detect the cell proliferation,Transwell test was used to measure the cell invasion and migration,and flow cytometry was used to detect the cell cycle and apoptosis.The correlation between CXCL5and tumor immune invasion level of LSCC was analyzed by ssGSEA,and CXCL5 co-expression gene network was constructed and analyzed for GO and KEGG enrichment.Results Compared with the adjacent tissues and the cells in control group,the expression of CXCL5 in LSCC tissues and cells increased,which was consistent with the analysis of TCGA database;Inhibition of CXCL5 expression in AMC-HN-8 cells inhibited the proliferation,invasion and migration of tumor cells,and promoted the apoptosis through inhibiting the cell cycle in G1 phase;The immune cell scores in DC,neutrophils,NK and TH17 cells were different.Conclusion CXCL5gene is highly expressed in LSCC tissues,which might be one of the targets of LSCC targeted therapy.

关 键 词：CXCL5 喉鳞状细胞癌 细胞生物学功能 调控 

分 类 号：R767[医药卫生—耳鼻咽喉科]