KIF14对肺腺癌A549细胞生物学功能的影响  

Effect of KIF14 on biological characteristics of lung adenocarcinoma A549 cells

作　　者：栾艳超[1] 彭海军[1] 杨倩 韩青松[1] 王明正[1] LUAN Yanchao;PENG Haijun;YANG Qian;HAN Qingsong;WANG Mingzheng(Hebei Provincial Lung Cancer Prevention and Treatment Research Center,Hebei Chest Hospital,Hebei Provincial Key Laboratory of Lung Disease,Hebei Shijiazhuang 050001,China)

机构地区：[1]河北省肺癌防治研究中心,河北省胸科医院,河北省肺病重点实验室,河北石家庄050001

出　　处：《现代肿瘤医学》2023年第14期2638-2646,共9页Journal of Modern Oncology

基　　金：河北省卫生健康委科研基金项目(编号:20231211);河北省财政厅省直医疗卫生机构老年病防治项目资助(编号:361013)。

摘　　要：目的:探讨驱动蛋白超家族蛋白14(KIF14)在肺腺癌(LUAD)中的表达情况及对LUAD细胞生物学功能的影响。方法:通过TCGA、GEO数据库分析KIF14 mRNA在LUAD中表达情况,采用Kaplan-Meier法分析KIF14表达与患者预后的关系。利用单样本基因集富集分析(ssGSEA)和TIMER数据库探索KIF14表达水平与肿瘤免疫浸润的关系。通过功能富集分析注释KIF14在肺腺癌中的生物学功能。进一步进行体外实验和免疫组织化学验证。通过实时荧光定量PCR(qPCR)和Western blot检测KIF14在LUAD细胞系中表达情况;构建KIF14干扰序列并将其转染入A549细胞中,将其分为NC组(转染KIF14阴性对照序列)、siKIF14组(转染靶向KIF14的siRNA序列)。Western blot检测细胞中KIF14沉默效果;CCK-8、平板克隆形成实验检测siKIF14对LUAD细胞增殖能力的影响;TUNEL凋亡实验检测siKIF14对LUAD细胞凋亡能力的影响;Transwell和划痕实验检测siKIF14对LUAD细胞侵袭和迁移能力的影响。结果:生物信息数据库分析显示,KIF14 mRNA在LUAD中高表达(P<0.05),且KIF14 mRNA高表达者累积总生存(OS)率较差(P<0.05);功能富集分析表明KIF14参与细胞分裂、细胞周期、DNA复制等过程。ssGSEA和TIMER分析显示KIF14表达与免疫细胞浸润呈负相关。LUAD组织及A549细胞中KIF14均高表达(P<0.05);沉默KIF14表达能够抑制A549细胞的增殖、侵袭和迁移能力,诱导细胞凋亡(P<0.05)。结论:KIF14可为肺腺癌诊治过程中的潜在靶点,它可能通过调控细胞增殖、迁移及侵袭等特性影响肺腺癌的发生、发展。Objective:To investigate the expression of kinesin superfamily protein 14(KIF14)in lung adenocarcinoma(LUAD)and its effect on the biological function of LUAD cells.Methods:The expression of KIF14 mRNA in LUAD was analyzed by TCGA and GEO databases.Kaplan-Meier method was used to analyze the relationship between KIF14 expression and the prognosis of patients.Single sample gene set enrichment analysis(ssGSEA)and TIMER database were used to explore the relationship between KIF14 expression level and tumor immune infiltration.The biological function of KIF14 in lung adenocarcinoma was annotated by functional enrichment analysis.Further in vitro experiments and immunohistochemistry were performed to verify the results.The expression of KIF14 in LUAD tissues and cell lines was detected by quantitative real-time PCR(qPCR)and Western blot.The KIF14 interference sequence was constructed and transfected into A549 cells,which were divided into NC group(transfected with KIF14 negative control sequence)and siKIF14 group(transfected with siRNA sequence targeting KIF14).The silencing effect of KIF14 in cells was detected by Western blot.CCK-8 and plate clone formation assay were used to detect the effect of siKIF14 on the proliferation of LUAD cells.TUNEL assay was used to detect the effect of siKIF14 on the apoptosis of LUAD cells.Transwell and scratch assays were used to detect the effect of siKIF14 on the invasion and migration of LUAD cells.Results:Bioinformatics database analysis showed that KIF14 mRNA was highly expressed in LUAD(P<0.05),and the cumulative overall survival(OS)rate of patients with high KIF14 mRNA expression was poor(P<0.05).Functional enrichment analysis showed that KIF14 was involved in cell division,cell cycle,DNA replication and other processes.ssGSEA and TIMER analysis showed a negative correlation between KIF14 expression and immune cell infiltration.KIF14 was highly expressed in LUAD tissues and A549 cells(P<0.05).Silencing KIF14 expression inhibited the proliferation,invasion and migration of A549

关 键 词：肺肿瘤 腺癌 细胞增殖 细胞凋亡 细胞运动 驱动蛋白超家族蛋白14 

分 类 号：R734.2[医药卫生—肿瘤]