背景:ASB6,E3泛素连接酶,通过泛素-蛋白酶体途径介导其底物蛋白的蛋白酶体降解。据报道,ASB6在几种生物过程中发挥重要作用,包括肿瘤干性和内质网应激。然而,ASB6在结直肠癌中的潜在作用和机制,特别是其与免疫浸润水平及其预后意义的关联,还有待充分阐明。方法:我们通过LASSO惩罚的Cox回归确定了CRC患者的关键预后基因,单变量和多变量Cox回归分析。随后,我们综合分析了hub基因的预后价值,并构建了预后列线图。最后,我们通过免疫沉淀-质谱(IP-MS)鉴定了ASB6相互作用蛋白,构建了蛋白-蛋白相互作用(PPI)网络,并进行了途径富集分析,以探索ASB6的潜在机制.同时,我们通过体外细胞实验评估了ASB6在CRC细胞中的功能。结果:我们将ASB6鉴定为CRC的hub基因。ASB6在CRC中高表达,ASB6高表达的患者无病间期(DFI)较差,疾病特异性生存率(DSS),总生存率(OS),和无进展间隔(PFI)。相关性分析显示ASB6的表达与淋巴结浸润和远端转移呈正相关。ASB6的过表达增强了CRC细胞的迁移能力。多因素Cox回归分析显示ASB6是影响CRCOS和DSS的独立预后因素。基于多变量分析结果构建的列线图模型具有较好的预测效果,OS和DSS的C指数为0.811和0.934,分别。此外,免疫浸润水平分析显示ASB6表达与M2型巨噬细胞浸润水平呈正相关,ASB6和M2型巨噬细胞水平高的患者预后较差.此外,通过IP-MS鉴定的ASB6相互作用蛋白的途径富集分析表明,ASB6可能通过对未折叠蛋白途径的反应和内质网途径的蛋白质加工发挥关键作用。结论:ASB6在CRC组织中显著上调,是影响CRC患者预后的危险因素。ASB6增强CRC细胞的迁移能力。因此,ASB6可能是CRC患者的独立预后生物标志物和潜在治疗靶标。
Background: ASB6, an E3 ubiquitin ligase, mediates the proteasomal degradation of its substrate proteins via the ubiquitin-proteasome pathway. ASB6 has been reported to play significant roles in several biological processes, including tumor stemness and endoplasmic reticulum stress. However, the underlying role and mechanism of ASB6 in colorectal cancer, particularly its association with immune infiltration levels and its prognostic significance, remain to be fully elucidated. Methods: We identified key prognostic genes in CRC patients through LASSO-penalized Cox regression, Univariate and Multivariate Cox regression analyses. Subsequently, we comprehensively analyzed the prognostic value of hub genes and constructed a prognostic nomogram. Finally, we identified ASB6 interacting proteins through immunoprecipitation-mass spectrometry (IP-MS) and constructed protein-protein interaction (PPI) networks and performed pathway enrichment analysis to explore the potential mechanisms of ASB6. Meanwhile, we evaluated the functions of ASB6 in CRC cells through in vitro cell experiments. Results: We identified ASB6 as a hub gene in CRC. ASB6 was highly expressed in CRC, and patients with high ASB6 expression had worse Disease-Free Interval (DFI), Disease-Specific Survival (DSS), Overall Survival (OS), and Progression-Free Interval (PFI). Correlation analysis showed that ASB6 expression were positively correlated with lymph node invasion and distal metastasis. Overexpression of ASB6 enhanced the migration ability of CRC cells. Multivariate Cox regression analysis revealed that ASB6 was an independent prognostic factor for OS and DSS in CRC. The nomogram model constructed based on multivariate analysis results had good predictive effects, with C-indexes of 0.811 and 0.934 for OS and DSS, respectively. Furthermore, analysis of immune infiltration levels showed that ASB6 expression were positively correlated with M2-type macrophage infiltration levels in CRC, and patients with high levels of both ASB6 and M2-type macrophages had a worse prognosis. Furthermore, pathway enrichment analysis of ASB6 interacting proteins identified by IP-MS suggested that ASB6 may play a crucial role through the response to unfolded protein pathway and protein processing in the endoplasmic reticulum pathway. Conclusions: ASB6 is significantly upregulated in CRC tissues and is a risk factor for prognosis in CRC patients. ASB6 enhances the migration ability of CRC cells. Therefore, ASB6 may be an independent prognostic biomarker and potential therapeutic target for CRC patients.