转录因子HOXC13通过上调PCNA表达促进喉鳞状细胞癌AMC-HN-8细胞的恶性生物学行为  

Transcription factor HOXC13 promotes malignant biological behaviors of laryngeal squamous cell carcinoma by upregulating PCNA expression

作　　者：兰利利[1] 牛云峰 胡国斌 刘猛[1] 徐玉茹[1] 王晶田[1] LAN Lili;NIU Yunfeng;HU Guobin;LIU Meng;XU Yuru;WANG Jingtian(Department of Otorhinolaryngology and Head and Neck Surgery,No.4 Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China;Department of Pathology,the 980th Hospital of the Joint Logistics Support Force,Shijiazhuang 050082,Hebei,China)

机构地区：[1]河北医科大学第四医院耳鼻咽喉头颈外科,河北石家庄050011 [2]联勤保障部队第九八〇医院病理科,河北石家庄050082

出　　处：《中国肿瘤生物治疗杂志》2024年第6期558-565,共8页Chinese Journal of Cancer Biotherapy

基　　金：河北省卫生健康委医学科学研究课题计划(No.20230907)。

摘　　要：目的:本研究旨在探讨转录因子HOXC13在喉鳞状细胞癌(LSCC)中的表达、功能及可能的调控机制。方法:常规培养LSCC细胞,将其分为sh-NC组、sh-HOXC13组、pcDNA3.1-NC组、pcDNA3.1-HOXC13组、pcDNA3.1-PCNA组和sh-HOXC13+pcDNA3.1-PCNA组,用转染试剂将相应核酸和质粒转染各组细胞。用数据库数据分析HOXC13mRNA在LSCC组织中的表达;收集2019年1月至2022年12月在联勤保障部队第九八〇医院耳鼻咽喉头颈外科手术切除的62对LSCC组织及配对的癌旁组织,免疫组织化学法检测中国人LSCC组织中HOXC13蛋白的表达,qPCR检测中国人LSCC组织、癌旁组织以及各组细胞中HOXC13和PCNAmRNA的表达,MTS法检测各组AMC-HN-8细胞的增殖能力,平板克隆实验检测各组AMC-HN-8细胞的克隆形成能力,Transwell小室实验检测各组AMC-HN-8细胞的迁移和侵袭能力,双萤光素酶报告基因实验和染色质免疫沉淀技术(ChIP)验证HOXC13与PCNA之间的结合关系。结果:HOXC13和PCNA在LSCC组织和细胞中均呈高表达(P<0.05或P<0.01)且两者的表达水平呈正相关(P<0.01),HOXC13的表达水平与TNM分期有明显关联(P<0.01)。敲减HOXC13可明显抑制AMC-HN-8细胞的增殖、迁移和侵袭能力(均P<0.01),过表达HOXC13则促进TU686细胞的增殖、迁移和侵袭能力(均P<0.01)。HOXC13可与PCNA启动子区结合并调控其转录。敲低PCNA可部分逆转HOXC13对AMC-HN-8细胞的恶性生物学行为的促进作用(均P<0.01)。结论:HOXC13通过上调PCNA促进LSCC细胞的恶性生物学行为,HOXC13是LSSC临床诊断和治疗的潜在靶点。Objective:To investigate the expression,function and possible regulatory mechanisms of transcription factor HOXC13 in laryngeal squamous cell carcinoma(LSCC).Methods:LSCC cells were routinely cultured and divided into sh-NC group,sh-HOXC13 group,pcDNA3.1-NC group,pcDNA3.1-HOXC13 group,pcDNA3.1-PCNA group,and sh-HOXC13+pcDNA3.1-PCNA group,and the corresponding nucleic acids and plasmids were transfected into each group of cells with transfection reagents.GEO Database data were used to analyze the expression of HOXC13 mRNA in LSCC tissues.Sixty-two pairs of LSCC tissues and paired adjacent tissues that were surgically removed in the 980th Hospital of the Joint Logistics Support Force between January 2019 and December 2022 were collected.The expression of HOXC13 protein in Chinese LSCC tissues were detected by immunohistochemistry.The expressions of HOXC13 and PCNA mRNA in Chinese LSCC tissues,adjacent non-cancerous tissues,and various cell lines were detected by qPCR.The proliferation ability of AMC-HN-8 cells in different groups was detected by the MTS method.Plate cloning assay was used to detect the clone formation ability of transfected AMC-HN-8 cells in each group.The migration and invasion abilities of AMC-HN-8 cells in each group were determined by transwell chamber experiments.The binding relationship between HOXC13 and PCNA was verified by dual luciferase reporter gene assays and chromatin immunoprecipitation(ChIP)techniques.Results:HOXC13 and PCNA were highly expressed in LSCC tissues and cells(P<0.05 or P<0.01).Their expression levels were positively correlated(P<0.01),and the expression level of HOXC13 was significantly correlated with TNM stage(P<0.01).Knockdown of HOXC13 significantly inhibited the proliferation,migration and invasion abilities of AMC-HN-8 cells(all P<0.01),and overexpression of HOXC13 promoted the proliferation,migration and invasion abilities of TU686 cancer cells(all P<0.01).HOXC13 could bind to the promoter region of PCNA and regulate its transcription.Knockdown of PCNA partially

关 键 词：转录因子 HOXC13 喉鳞状细胞癌 AMC-HN-8细胞 增殖 侵袭 迁移 PCNA 

分 类 号：R739.65[医药卫生—肿瘤]