TRAP1在结肠癌组织中的表达与病理特征和患者预后的关系及其可能的机制

Expression of TRAP1 in colon cancer tissues and its relationship with clinicopathological features and patient prognosis and the possible molecular mechanisms

目的:探讨肿瘤坏死因子受体相关蛋白1(TRAP1)在结肠癌组织和细胞中的表达及其与临床病理特征和患者预后的关系和相关分子机制。方法:通过TCGA和GEO数据全面分析TRAP1在结肠癌中的表达及其与临床病理特征和患者预后的关系,选取2020年10月至2021年03月间在山西医科大学第一医院手术切除的10例结肠癌组织及相应癌旁组织标本,用IHC染色法检测中国人结肠癌组织中TRAP1的表达进行验证,运行R包(survival和survminer)进行Kaplan-Meier生存分析;在线分析TRAP1蛋白的信号肽及穿膜结构域,通过基因富集分析软件进行GO分析和KEGG分析。培养结肠癌SW480和SW620细胞,将si-NC和si-TRAP1转染结肠癌细胞,实验分为空白对照组、si-NC组和si-TRAP1组,采用qPCR法检测转染后各组结肠癌细胞中TRAP1的表达,FCM检测转染后各组细胞的细胞周期和凋亡情况。结果:与癌旁组织比较,TRAP1在结肠癌组织中呈高表达(P<0.01),TRAP1表达水平与淋巴结转移有关联(P<0.05),TRAP1高表达组结肠癌患者5年OS率较低(P<0.05)。TRAP1蛋白属于细胞质蛋白,功能富集结果显示TRAP1及其相关分子与细胞周期、核糖体生物发生等信号通路有关(均P<0.01),TRAP1高表达组的结肠癌代谢重编程基因簇和线粒体蛋白输入基因簇水平升高(均P<0.01)。敲减TRAP1后,结肠癌细胞周期阻滞于G1期,细胞凋亡水平显著升高(均P<0.01)。结论:TRAP1在结肠癌组织中呈高表达,且与患者淋巴结转移和低OS率相关联,敲减TRAP1可阻滞结肠癌细胞周期并促进其凋亡。

Objective:To investigate the expression of tumor necrosis factor receptor associated protein 1(TRAP1)in colon cancer tissues and its relationship with clinicopathological features and patient prognosis, as well as the related molecular mechanisms.Methods:The expression of TRAP1 in colon cancer and its relationship with clinicopathological features and patient prognosis were comprehensively analyzed by TCGA and GEO databases. In addition, 10 pairs of colon cancer tissues and corresponding paracancerous tissues surgically resected at the First Hospital of Shanxi Medical University between October 2020 and March 2021 were selected, and the expression of TRAP1 in colon cancer tissues of Chinese was detected by IHC staining for validation. Kaplan-Meier survival analysis was performed by running R package(survivor and survminer);The signal peptide and membrane piercing domain of TRAP1protein were analyzed by online software, and GO analysis and KEGG analysis were carried out by gene enrichment analysis software.Colon cancer SW480 and SW620 cells were cultured and transfected with si-NC or si-TRAP1, and divided into blank control group,si-NC group and si-TRAP1 group. The TRAP1 expression in colon cancer cells of each group after transfection was detected by qPCR,and the cell cycle and apoptosis of the transfected cells were detected by FCM technique. Results:TRAP1 was highly expressed in colon cancer tissues compared with paracancerous tissues(P<0.01). The expression level of TRAP1 was correlated with lymph node metastasis(P<0.05), and the 5-year overall survival(OS)rate of colon cancer patients with high TRAP1 expression was lower than those with low TRAP1 expression(P<0.05). TRAP1 protein is a cytoplasmic protein, and functional enrichment results showed that TRAP1 and its related molecules are associated with signaling pathways such as cell cycle and ribosome biogenesis(all P<0.01). GSEA enrichment results showed that the levels of colon cancer metabolic reprogramming gene cluster and mitochondrial protein input gene cluster were elevated in the high TRAP1 expression group(all P<0.01). Knockdown of TRAP1 resulted in cell cycle arrest in G1 phase with significantly elevated level of apoptosis of colon cancer cells(all P<0.01). Conclusions:TRAP1 is highly expressed in colon cancer tissues and correlated with lymph node metastasis and dismal OS rate in patients. Knockdown of TRAP1 can block cell cycle and promote apoptosis in colon cancer cells.