泛素C端水解酶37通过去泛素化巨噬细胞迁移抑制因子促进结肠癌细胞增殖

Ubiquitin C-terminal hydrolase 37 promotes the proliferation of colon cancer cells by de-ubiquitinating MIF

目的检测UCH37在结肠癌中的表达,并分析其与患者的临床病理特征的相关性,及其对结肠癌细胞增殖和凋亡的影响及分子机制。方法利用免疫组化检测结肠癌组织中UCH37蛋白的表达情况,分析UCH37与结肠癌患者临床病理特征的相关性,qPCR、CCK-8、流式细胞术和免疫共沉淀检测UCH37对结肠癌细胞增殖、凋亡及MIF的去泛素化水平的影响。结果 UCH37蛋白在结肠癌组织中的阳性率高于癌旁组织,其表达水平与患者TNM分期及核分化水平呈正相关;结肠癌细胞中UCH37的表达水平显著高于正常结肠内皮细胞;转染UCH37 siRNA组细胞增殖能力低于转染NC组,而凋亡细胞比例高于转染NC组;感染UCH37慢病毒组增殖能力高于感染空载体组,而凋亡细胞比例低于感染空载体组。其机制为UCH37在结肠癌细胞中去泛素化MIF。结论 UCH37在结肠癌中表达升高,并通过去泛素化MIF促进结肠癌细胞增殖。

Objective To explore the expression and cilinical significance of UCH37 in colon cancer, and detect the effects and underlying mechanism of UCH37 on cell proliferation and cell apoptosis of colon cancer cell lines. Methods Expressions of UCH37 were analyzed by immunohistochemistry in colon cancer patients. Spearman′s rank test was conducted to analyze the clinical relevance of UCH37 in colon cancer. QPCR was conducted to detect the expression of UPS39 in colon cancer cell lines and NCM460 cells.CCK-8,flow cytometry, co-immunoprecipitation were conducted to detect the effects of UCH37 on cell proliferation,cell apoptosis. And ubiquitinated level of MIF. Results Compare to adjacent tissues,immunohistochemistry and chi square analysis revealed that the positive rate of UCH37 was upregulated in colon cancer tissues.Spearman rank correlation showed that positive UCH37 expression was significantly associated with TNM stage and cell differentiation of colon cancer patients. CCK-8 results showed cell proliferation in colon cancer cells transfected with UCH37 NC was promoted and cell apoptosis in colon cancer cells transfected with UCH37 NC was inhibited compared with cells transfected UCH37 siRNAs;furthermore,compared to cells transfected lentiviral vector carrying pLenti6.3.cell proliferation in colon cancer cells transfected with lentiviral vector carrying UCH37 was promoted and cell apoptosis in colon can-cer cells transfected with lentiviral vector carrying UCH37 was inhibited.Co-IP showed MIF could be deubiquitinat-ed by UCH37 in colon cancer cells. Conclusion UCH37 expression is upregulated in colon cancer,and UCH37 could promote cell proliferation of colon cancer cells by deubiquitinating MIF.