UCHL1通过调控肿瘤微环境糖酵解代谢促进肺腺癌细胞增殖和转移

UCHL1 promotes the proliferation and metastasis of lung cancer cells by regulatingglycolytic metabolism in tumor microenvironment

目的:探讨泛素羧基末端水解酶L1(ubiquitin C-terminal hydrolase-L1,UCHL1)通过调控缺氧诱导因子1(hypoxia-inducible factor 1,HIF-1)介导的代谢重编程促进肺腺癌细胞的增殖和转移。方法:免疫组化和实时定量聚合酶链反应检测肺腺癌组织和细胞中UCHL1和HIF-1的表达;Kaplan-Meier Plotter和UALCAN数据库分析了UCHL1和HIF-1的表达量与患者生存期的关联,并进一步分析了二者在不同临床病理等级以及淋巴转移患者肿瘤组织中的表达;克隆形成、迁移和侵袭评估肺腺癌HCC4006细胞转染UCHL1 siRNA后恶性生物学行为变化;Western Blot检测沉默UCHL1后肿瘤细胞糖酵解信号通路IL-6/STAT3标志分子的表达。结果:免疫组化、实时定量聚合酶链反应以及相关性分析,结果显示肺腺癌组织和细胞中UCHL1和HIF-1的表达较癌旁组织和正常人支气管上皮细胞中显著增加,且二者表达水平呈正相关(P<0.01),与Oncomine数据库中UCHL1和HIF-1的表达趋势相一致。生信分析结果显示,UCHL1和HIF-1异常高表达与肺腺癌患者的生存期呈负相关、而与患者的病理等级和淋巴结转移呈正相关(P<0.01)。转染UCHL1 siRNA的肺腺癌HCC4006细胞的克隆形成、迁移和侵袭能力较对照组细胞显著降低,同时细胞的耗氧量显著增加,并抑制细胞中LDH活性和LD分泌(P<0.01)。Western Blot结果显示,沉默UCHL1可抑制HIF-1表达,并降低缺氧介导的肿瘤细胞糖酵解信号通路IL-6/STAT3标志分子的表达(P<0.01)。结论:肺腺癌细胞通过UCHL1-HIF-1轴介导的代谢重编程获得较强的增殖和转移表型,为靶向该基因网络的治疗提供了理论依据。

Objective:To explore the effect of ubiquitin C-terminal hydrolase-L1(UCHL1)on the proliferation and metastasis of lung cancer cells by regulating hypoxia-inducible factor 1(HIF-1)mediated metabolic reprogramming.Methods:Immunohistochemistry and real-time quantitative polymerase chain reaction were used to detect the expression of UCHL1 and HIF-1 in lung cancer tissues and cells.Kaplan-Meier Plotter and UALCAN databases analyzed the correlation between the expression levels of UCHL1 and HIF-1 and patient survival,and further analyzed their expression in tumor tissues of patients with different clinical pathological grades and lymphatic metastasis.Clone formation,migration,and invasion assessment of malignant biological behavior changes in lung cancer HCC4006 cells transfected with UCHL1 siRNA.Western Blot detection of the expression of IL-6/STAT3 markers in tumor cells after silencing UCHL1.Results:Immunohistochemistry,real-time quantitative polymerase chain reaction,and correlation analysis,the results showed that the expressions of UCHL1 and HIF-1 in lung cancer tissues and cells were significantly increased compared with those in adjacent tissues and normal bronchial epithelial cells,and the expression levels of the two were positively correlated ( P < 0. 01) , and these results were consistent with the expression trend ofUCHL1 and HIF - 1 in Oncomine database. The results of bioinformatics analysis showed that the abnormally high expressionof UCHL1 and HIF - 1 was negatively correlated with the survival time of lung cancer patients,but positivelycorrelated with the pathological grade and lymph node metastasis of patients ( P < 0. 01) . The ability of colony formation,migration, and invasion of lung cancer HCC4006 cells transfected with UCHL1 siRNA was significantly reducedcompared with the control cells, the oxygen consumption of the cells was significantly increased, and the LDH activityand LD secretion were inhibited ( P < 0. 01) . Western Blot results showed that silencing UCHL1 inhibited HIF - 1 expressionand decreased the expression of tumor cell glycolysis IL - 6 /STAT3 signaling pathway biomarkers mediatedby hypoxia ( P < 0. 01) . Conclusion: Lung cancer cells acquire strong proliferative and metastatic phenotypes throughmetabolic reprogramming mediated by the UCHL1 - HIF - 1 axis,providing a rationale for therapies targeting thisgene network.