CD2AP在胰腺癌中的预后价值及对细胞增殖、侵袭的影响

Prognostic value of CD2AP in pancreatic cancer and its effects on cell proliferation and invasion

摘要:

目的 探讨CD2相关蛋白（CD2AP）在胰腺癌中的预后价值及对细胞增殖、侵袭的影响。方法 通过肿瘤基因组图谱（TCGA）、组织-基因型表达数据库及基因表达数据库中的GSE62452、GSE183795数据集分析CD2AP在胰腺癌中的表达及预后意义；基于TCGA中胰腺癌表达谱数据结合基因富集分析算法分析CD2AP与代谢通路的相关性，使用TIMER 2.0软件分析CD2AP与胰腺癌突变基因、免疫细胞浸润丰度的关系。采用EdU免疫荧光染色、Transwell实验及划痕实验研究CD2AP对胰腺癌细胞增殖、侵袭和迁移的影响。结果 TCGA、GSE62452、GSE183795数据集肿瘤组织中CD2AP表达量高于癌旁组织（P <0.05）。CD2AP高表达组预后均较CD2AP低表达组差（P <0.05）。在CD2AP高表达组中，显著上调的有皮质激素合成、卟啉代谢及亮氨酸和异亮氨酸合成信号通路，而钙离子、神经活性配体-受体相互作用信号通路则显著下调。突变型TP53、KRAS、CDKN2A基因的CD2AP表达量均高于野生型（P <0.05）。突变型与野生型SMAD4基因的CD2AP表达量比较，差异无统计学意义（P >0.05）。Spearman相关性分析结果显示，CD2AP的表达与CD8⁺T细胞、树突状细胞、B细胞和中性粒细胞浸润丰度呈正相关（r_s =0.363、0.280、0.363和0.237，均P <0.05），而与CD4⁺T细胞、巨噬细胞浸润丰度无相关性（r_s =-0.117和0.101，均P >0.05）。CD2AP-OE组CD2AP相对表达量高于Control组（P <0.05），CD2AP-siRNA组CD2AP相对表达量低于NC-siRNA组（P <0.05）。CD2AP-OE组细胞增殖率较Control组高（P <0.05），CD2AP-siRNA组细胞增值率较NC-siRNA组低（P <0.05）。CD2AP-OE组细胞侵袭数较Control组多（P <0.05），CD2AP-siRNA组细胞侵袭数较NC-siRNA组少（P <0.05）。CD2AP-OE组细胞迁移率较Control组高（P <0.05），CD2AP-siRNA组较NC-siRNA组低（P <0.05）。结论 CD2AP表达水平可作为评估胰腺癌预后的肿瘤标志物，是胰腺癌治疗的潜在靶点。

Abstract:

Objective To explore the prognostic value of CD2-associated protein (CD2AP) in pancreatic cancer and its effects on cell proliferation and invasion.Methods The expression and prognostic value of CD2AP in pancreatic cancer were analyzed using The Cancer Genome Atlas (TCGA) database, Genotype-Tissue Expression (GTEx) database and GSE62452 and GSE183795 datasets from the Gene Expression Omnibus (GEO) database. The correlation between CD2AP and metabolic pathways was analyzed with the GSEA algorithm based on the pancreatic cancer expression profiling data in the TCGA database. The associations between CD2AP and mutant genes in pancreatic cancer, as well as the infiltration of immune cells, were analyzed by the TIMER 2.0 web tool. The EdU assay, Transwell assay, and scratch assay were performed to determine the effects of CD2AP on proliferation, invasion and migration of pancreatic cancer cells.Results The expression of CD2AP was found to be up-regulated in pancreatic cancer tissues compared to that in adjacent tissues based on the TCGA database and the GSE62452 and GSE183795 datasets (P < 0.05). The prognosis of the CD2AP high-expression group was poorer than that of the CD2AP low-expression group (P < 0.05). In the CD2AP high-expression group, signaling pathways such as corticosteroid synthesis, porphyrin metabolism, and leucine and isoleucine synthesis were significantly up-regulated, whereas the calcium signaling pathway and the neuroactive ligand-receptor interaction signaling pathway were significantly down-regulated (P < 0.05). Upregulation of CD2AP expression was observed in patients with TP53, KRAS and CDKN2A mutations (P < 0.05). There was no difference in the expression of CD2AP between those with mutant and wild-type SMAD4 (P > 0.05). Spearman correlation analysis revealed that the expression of CD2AP was positively correlated with the abundance of infiltrated immune cells including CD8⁺T cells, dendritic cells, B cells and neutrophils (r_s = 0.363, 0.280, 0.363 and 0.237, all P < 0.05) but not correlated with the abundance of the infiltration of CD4⁺T cells and macrophages (r_s = -0.117 and 0.101, both P < 0.05). The relative expression of CD2AP in the CD2AP-OE group was higher than that in the control group (P < 0.05), while that in the CD2AP-siRNA group was lower than that in the NC-siRNA group (P < 0.05). The cell proliferation rate of the CD2AP-OE group was higher than that of the control group (P < 0.05), and that of the CD2AP-siRNA group was lower than that of the NC-siRNA group (P < 0.05). The number of invaded cells in the CD2AP-OE group was higher than that in the control group (P < 0.05), and that in the CD2AP-siRNA group was lower than that in the NC-siRNA (P < 0.05). The cell migration rate in the CD2AP-OE group was higher than that in the control group (P < 0.05), and that in the CD2AP-siRNA group was lower than that in the NC-siRNA (P < 0.05).Conclusions CD2AP can be used as a tumor marker to assess the prognosis of patients with pancreatic cancer and a potential target for the treatment of pancreatic cancer.