摘要
目的观察着丝粒蛋白F(CENPF) mRNA在肺腺癌细胞系中的表达变化,分析CENPF mRNA在肺腺癌组织中的表达变化并探讨其相关调控信号通路。方法①采用实时定量PCR法检测肺正常上皮细胞系BEAS-2B、肺腺癌细胞系(A549、NCI-H1650、NCI-H23、NCI-H1975) CENPF mRNA。②采用癌症基因组图谱(TCGA)的肺腺癌数据集分析CENPF mRNA表达与肺腺癌临床病理参数、预后的关系,Cox回归模型分析肺腺癌患者预后的影响因素。③采用基因集富集分析(GSEA)法分析受CENPF调控的肺腺癌相关信号通路。结果①A549、NCI-H1650、NCI-H23、NCI-H1975、BEAS-2B细胞CENPF mRNA相对表达量分别为4. 333±0. 271、3. 193±0. 188、2. 770±0. 091、5. 491±0. 249、1. 006±0. 071,A549、NCI-H1650、NCI-H23、NCI-H1975细胞CENPF mRNA相对表达量均高于BEAS-2B(P均<0. 05)。②肺腺癌、癌旁正常组织CENPF mRNA的相对表达量分别为9. 632±0. 059、6. 472±0. 093,二者比较,P <0. 001。CENPF mRNA表达与肺腺癌患者的性别、年龄、T分期、N分期及TNM分期相关(P均<0. 05)。生存分析结果显示,与CENPF mRNA低表达患者比较,CENPF mRNA高表达的肺腺癌患者的预后差(P=0. 001)。CENPF mRNA表达、临床TNM分期(HR分别为1. 667,1. 414; P均<0. 01)是影响肺腺癌患者预后的因素。③与CENPF mRNA高表达有关的肺腺癌调控信号通路有G2M检查点(P <0. 001,FDR <0. 001)、有丝分裂纺锤体(P <0. 001,FDR <0. 001)、E2F靶基因(P <0. 001,FDR <0. 001)、MYC靶基因(P <0. 001,FDR=0. 001)、m TOR信号通路(P=0. 002,FDR=0. 005)、精子形成(P=0. 002,FDR=0. 009)、未折叠蛋白反应(P=0. 002,FDR=0. 020)及PI3K/Akt/m TOR信号通路(P=0. 015,FDR=0. 117)等。结论肺腺癌组织及细胞系中存在CENPF高表达,CENPF高表达与肺腺癌患者的恶性病理特征及不良预后密切相关。CENPF mRNA高表达是影响肺腺癌患者预后的独立危险因素。CENPF可能通过调节G2M检查点、有丝分裂纺锤体、E2F靶基因、MYC靶基因、m TOR信号通路、精子形成、未折叠蛋白反应及PI3K/Akt/m TOR信号通路在肺腺癌的发生、发展中起重要作用。
Objective To observe the expression changes of centromere protein F( CENPF) mRNA in the lung adenocarcinoma cell lines,and to analyze the expression changes of CENPF mRNA in the lung adenocarcinoma tissues and to explore its related regulatory signaling pathways. Methods ① The real-time quantitative PCR was used to detect the CENPF mRNA in the normal lung epithelial cell line BEAS-2 B and lung adenocarcinoma cell lines( A549,NCI-H1650,NCIH23,and NCI-H1975). ② The relationships of CENPF mRNA with clinicopathological characteristics and prognosis of lung adenocarcinoma were analyzed by the lung adenocarcinoma dataset of Cancer Genome Atlas( TCGA). Cox regression model was used to analyze the prognostic factors of lung adenocarcinoma patients. ③ Gene Set Enrichment Analysis( GSEA) was used to analyze the signal pathways involved in lung adenocarcinoma regulated by CENPF. Results ① The relative expression of CENPF mRNA in the A549,NCI-H1650,NCI-H23,NCI-H1975,and BEAS-2 B cells were 4. 333 ±0. 271,3. 193 ± 0. 188,2. 770 ± 0. 091,5. 491 ± 0. 249,and 1. 006 ± 0. 071,respectively. The relative expression of CENPF mRNA in the A549,H1650,NCI-H23,and NCI-H1975 cells was higher than that in the BEAS-2 B cells( P <0. 05). ② The relative expression levels of CENPF mRNA in the lung adenocarcinoma and adjacent normal tissues were9. 632 ± 0. 059 and 6. 472 ± 0. 093,with significant difference( P < 0. 001). CENPF mRNA expression was related to gender,age,T stage,N stage,and TNM stage( all P < 0. 05). Survival analysis showed that patients with high CENPF mRNA expression had poorer prognosis compared with patients with low CENPF mRNA expression( P = 0. 001). Cox regression model analysis showed that CENPF mRNA expression and TNM staging were independent risk factors for the prognosis of lung adenocarcinoma patients( HR = 1. 667 and 1. 414,respectively,both P < 0. 01). ③ The lung adenocarcinoma regulatory signaling pathway associated with high expression of CENPF mRNA included G2 M checkpoint( P < 0. 001,FDR <0. 001),mitotic spindle( P < 0. 001,FDR < 0. 001),E2 F target gene( P < 0. 001,FDR < 0. 001),MYC target gene( P< 0. 001,FDR = 0. 001),m TOR signaling pathway( P = 0. 002,FDR = 0. 005),spermatogenesis( P = 0. 002,FDR =0. 009),unfolded protein response( P = 0. 002,FDR = 0. 020),and PI3 K/Akt/m TOR signaling pathway( P = 0. 015,FDR = 0. 117). Conclusions CENPF mRNA is highly expressed in the lung adenocarcinoma tissues and cell lines. The high expression of CENPF mRNA is closely related to the clinicopathological characteristics and poor prognosis of lung adenocarcinoma patients. High expression of CENPF mRNA is an independent risk factor for the prognosis of lung adenocarcinoma patients. CENPF mRNA may play an important role in the development and progression of lung adenocarcinoma by regulating G2 M checkpoint,mitotic spindle,E2 F target gene,MYC target gene,m TOR signaling pathway,spermatogenesis,unfolded protein response,and PI3 K/Akt/m TOR signaling pathway.
引文
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