CENPF通过上调ACKR3/CXCR7促进非小细胞肺癌EMT的研究

顾彤; 姜瑜珩; 丁姝; 陈炜; 罗超; 于伟勇; 陈小飞

doi:10.3971/j.issn.1000-8578.2022.22.0322

CENPF通过上调ACKR3/CXCR7促进非小细胞肺癌EMT的研究

CENPF Promotes EMT in Non-small Cell Lung Cancer by Up-regulating ACKR3/CXCR7

摘要

摘要:
目的探讨CENPF在非小细胞肺腺癌（LUAD）中的表达与患者临床预后的关系及其对肺腺癌细胞转移能力的影响。
方法公共数据库分析CENPF在LUAD中的表达及其与患者预后的关系。免疫组织化学染色验证CENPF LUAD在组织芯片中的表达，Kaplan-Meier分析CENPF表达与肺腺癌患者预后的关系；Cox生存风险比例回归模型分析影响患者生存的因素；卡方分析CENPF表达与患者临床病理分期及分级的关系。慢病毒敲除NCI-H2126细胞中CENPF的表达，检测细胞增殖、侵袭及迁移能力的变化。RNA-seq检测CENPF敲除后细胞mRNA表达谱改变，生物信息学分析CENPF下游信号通路及靶基因，Western blot验证下游基因表达水平变化。
结果 CENPF在LUAD肿瘤组织中显著上调（P < 0.05），与病理分期显著相关（P=0.013），表达越高患者预后越差（P=0.01, P=0.027）。敲除CENPF表达后，细胞增殖、迁移及侵袭能力均显著降低（P < 0.01）；RNA-seq富集分析显示细胞趋化因子通路基因表达改变富集显著（P < 0.001）；聚类差异分析则表明，ACKR3/CXCR7及CDH2/N-cadherin显著下调，CDH1/E-cadherin则显著上调；Western blot结果证实，敲除CENPF后，ACKR3/CXCR7及N-cadherin显著下调，E-cadherin则显著上调。
结论 CENPF的表达与LUAD患者临床预后呈负相关，其通过ACKR3/CXCR7调控与EMT相关的N-cadherin及E-cadherin的表达促进EMT的发生。

Abstract:
Objective To investigate the relationship between the expression of CENPF in NSCLC adenocarcinoma (LUAD) and the clinical prognosis of patients and its effect on the metastasis of lung adenocarcinoma cells.
Methods The expression of CENPF in LUAD and its relationship with patient prognosis were analyzed by online bioinformatics. The expression of CENPF was verified by LUAD tissue microarray immunohistochemical staining. Kaplan-Meier analysis was performed to analyze the relationship between the expression of CENPF and the prognosis of patients with lung adenocarcinoma. Cox survival hazard ratio was used to analyze the factors affecting the survival of patients. Chi-square analysis was adopted to examine the relationship between CENPF expression and clinicopathological stage and grade of patients. The expression of CENPF in NCI-H2126 cells were knocked out by lentivirus, and then the proliferation, invasion, and migration abilities of the cells were detected. Changes in mRNA expression profiles after CENPF knockout were detected by RNA-seq. Bioinformatics analysis of downstream signaling pathways and the target genes of CENPF was also performed. Western blot was used to verify the target gene.
Results CENPF was significantly upregulated in LUAD tumor tissue (P < 0.05) and significantly correlated with pathological stage (P=0.013). The higher expression of CENPF, the worse the prognosis of patients (P=0.01, P=0.027). After the expression was CENPF of knocked out, the cell proliferation, migration, and invasion abilities significantly reduced (P < 0.01). The expression of chemokine pathway genes in cells was enriched significantly (P < 0.001). ACKR3/CXCR7 and CDH2/N-cadherin were significantly downregulated, whereas CDH1/E-cadherin was significantly upregulated. After CENPF was knocked out, ACKR3/CXCR7 and N-cadherin were significantly downregulated, whereas E-cadherin significantly increased.
Conclusion The expression of CENPF is negatively correlated with the clinical prognosis of patients with LUAD, and it promotes the occurrence of EMT by regulating the expression levels of N-cadherin and E-cadherin related to EMT through ACKR3/CXCR7.

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