摘要
目的探讨CENPF在肺鳞癌组织中的表达与患者临床预后关系及其对肺鳞癌细胞侵袭、转移能力的影响。方法收集90例肺鳞癌患者180个组织样本点,含配对癌组织和癌旁组织。生物信息学分析筛选公共数据库中与肺鳞癌预后相关基因CENPF。组织芯片免疫组化验证CENPF表达,Cox风险分析影响患者生存的病理因素,CENPF表达与患者预后、临床病理分期及分级的关系。敲减SK-MES-1细胞中CENPF表达,检测细胞增殖、侵袭及迁移能力;RNA-seq检测mRNA表达谱分析下游信号通路及靶基因;验证基因表达及细胞信号通路激活。结果数据库及临床样本组化结果均证实CEPNF在肺鳞癌细胞中表达上调(t=7.821,P<0.001),并与肺鳞癌患者预后相关(χ^(2)=4.09,P<0.001),且CENPF与肺鳞癌患者N分期相关(χ^(2)=7.869,P=0.005)。敲减CENPF表达后,细胞增殖(t=5.319,P<0.01)及迁移(t=13.72,P<0.001)和侵袭能力(t=7.555,P<0.001)显著增强;RNA-seq显示细胞黏附分子通路基因表达改变显著(P<0.001)。Western blot及qPCR表明敲减CENPF后SK-MES-1细胞中CLDN1(Claudin-1)及E-cadherin表达显著下调,而N-cadherin则显著上调(均P<0.05);Akt激活水平显著升高,且抑制Akt激活后CLDN1表达下调则被消除(均P<0.05)。结论CENPF的表达与肺鳞癌患者临床预后呈正相关,其通过抑制Akt信号通路的激活上调CLDN1的表达抑制肺鳞癌的转移。
Objective To investigate the CENPF expression in lung squamous cell carcinoma(LUSC)and its relationship to the clinical prognosis of patients,and to analyze the effect of CENPF on LUSC metastasis.Methods The 180 tissue sample points from 90 LUSC patients containing paired and para-cancer tissues.The CENPF gene,which has been associated with the progression of LUSC,was screened in public databases through bioinformatics analysis.Immune tissue micro-array results validated CENPF expression.Pathological factors affecting patient survival and the relationship between the CENPF expression and the prognosis of LUSC patients and clinicopathological staging and grading were analyzed by Cox risk mode.Gene expression and cell signaling pathway activation were verified in two ways.The first was by knockdown of CENPF expression in SK-MES-1 LUSC cells to detect cell proliferation,cell invasion,and cell migration.The second way was that the mRNA expression profile was revealed using RNA-seq to analyze downstream signaling pathways and target genes.Results Both public databases and immunohistochemical results of clinical samples confirmed the upregulation of CEPNF expression in LUSC(t=7.821,P<0.001).Cox risk analyses presented that the CENPF expression correlated with the prognosis of LUSC patients(χ^(2)=4.09,P<0.001)and the N staging(χ^(2)=7.869,P=0.005).Knockdown of CENPF expression significantly enhanced cancer cell proliferation(t=5.319,P<0.01),migration(t=13.72,P<0.001),and invasion(t=7.555,P<0.001).RNA-seq revealed significant alterations in gene expression of cell adhesion molecule pathway(P<0.001).After knockdown of CENPF,Western blot and qPCR revealed significant down-regulation of CLDN1(Claudin-1)and E-cadherin expressions,while N-cadherin was significantly upregulated in SK-MES-1 cells(P<0.05).Akt activation significantly increased,and inhibition of Akt activation eliminated the downregulation of CLDN1 expression(P<0.05).Conclusions The CENPF expression was positively correlated with the clinical prognosis of LUSC patients.CENPF inhibited LUSC metastasis by upregulating CLDN1 expression through the inhibited activation of the Akt signaling pathway.
作者
姜瑜珩
刘当云
丁姝
朱子薇
罗超
陈炜
陈小飞
Jiang Yuheng;Liu Dangyun;Ding Shu;Zhu Ziwei;Luo Chao;Chen Wei;Chen Xiaofei(Department of Oncology,Huai’an First Hospital Affiliated to Nanjing Medical University,Huaian 223300,China;Central Laboratory of Huai’an First Hospital Affiliated to Nanjing Medical University,Huaian 223300,China;Department of Respiratory and Critical Care Medicine,Huaian Clinical College of Xuzhou Medical University,Huaian 223300,China)
出处
《中华转移性肿瘤杂志》
2022年第4期317-325,共9页
Chinese Journal of Metastatic Cancer
基金
南京医科大学附属淮安第一医院高层次人才项目(YGRX201917)
淮安市免疫学重点实验室项目(HAP202002)
南京医科大学科技发展基金项目(NMUB20210141)
淮安市自然科学基金项目(HAB 201928、HAB 202204)
作者简介
通信作者:陈炜,Email:hayychw@njmu.edu.cn;通信作者:陈小飞,Email:hayylch@njmu.edu.cn;刘当云,对本文有同等贡献;姜瑜珩,对本文有同等贡献
