去泛素化酶泛素特异性肽酶14促进胰腺癌细胞的增殖和迁移能力

The Deubiquitinating Enzyme USP14 Promotes the Proliferation and Migration Ability of Pancreatic Cancer Cells

胰腺癌仍然是最致命的癌症之一,而且较少发现有效的治疗方案。虽然在许多肿瘤细胞,包括胰腺癌细胞中观察到泛素特异性蛋白酶14(ubiquitin specific peptidase 14,USP14)的过度表达,但它在胰腺癌中的确切作用仍未得到较好的阐明。我们研究了USP14在胰腺癌中的生物学功能及其分子机制。对癌症基因组图谱数据库(The Cancer Genome Atlas,TCGA)的分析显示USP14在胰腺癌组织中高度表达,进一步探究发现,其表达水平与患者的预后呈负相关。利用shRNAUSP14慢病毒建立了稳定的USP14敲低胰腺癌细胞系,通过CCK8、克隆形成实验、划痕和Transwell试验发现,USP14敲低抑制了胰腺癌细胞的增殖和迁移能力。在胰腺癌细胞SW1990和MIAPaCa2中蛋白质印迹结果表明,下调USP14的表达导致细胞周期蛋白D3(cyclin D3)水平下降,而过表达USP14会增加细胞周期蛋白D3表达水平。此外,免疫共沉淀证明,USP14与细胞周期蛋白D3相互作用,泛素化结果显示,USP14的过表达降低了细胞周期蛋白D3的泛素化水平。并且在SW1990胰腺癌细胞中,CRISPR/Cas9介导的USP14敲除导致细胞周期蛋白D3水平下降。这些发现表明,USP14通过与细胞周期蛋白D3的相互作用促进了胰腺癌细胞的增殖和迁移能力,凸显了USP14作为胰腺癌的潜在治疗靶点。

Pancreatic cancer remains one of the deadliest cancer types with few effective treatment options.While the overexpression of ubiquitin-specific protease 14(USP14)has been observed in many tumor cells,including pancreatic cancer cells,its precise role in pancreatic cancer is not well defined.Here,we investigated the biological function of USP14 in pancreatic cancer and its molecular mechanisms.Our analysis of the Cancer Genome Atlas database revealed that USP14 was highly expressed in pancreatic cancer tissues,and further investigation revealed that its expression level was negatively correlated with the prognosis of patients.In SW1990 and MIAPaCa2 pancreatic cancer cells,we established stable USP14-knockdown cell lines using the shRNA-USP14 lentivirus and found that USP14 knockdown inhibited the proliferation and migration ability of pancreatic cancer cells by CCK8,colony formation assay,wound-healing and Transwell assays.Western blotting analysis showed that downregulation of USP14 expression resulted in a decrease in CyclinD3 protein levels,while overexpression of USP14 increased the protein levels in SW1990 and MIAPaCa2 pancreatic cancer cells.Furthermore,co-immunoprecipitation demonstrated that USP14 interacts with CyclinD3 and ubiquitination assays show that overexpression of USP14 reduces the ubiquitination level of CyclinD3.Moreover,CRISPR/Cas9-mediated USP14 knock-out in SW1990 pancreatic cancer cells resulted in decreased CyclinD3 protein levels.These findings sug-gest that USP14 promotes the proliferation and migration ability of pancreatic cancer cells by interacting with CyclinD3,highlighting USP14 as a potential therapeutic target for pancreatic cancer.