ATP2A1影响结直肠癌增殖、迁移及免疫浸润的机制研究

ATP2A1 regulates proliferation and migration in colorectal cancer and serves as a predictor of patient prognosis and immune infiltration

目的:探究内质网Ca2+转运ATP酶亚型1(ATP2A1)参与影响结直肠癌(CRC)生物学行为和免疫浸润的机制。方法:采用转录组及免疫组化(IHC)分析ATP2A1在结直肠癌肿瘤细中表达与结直肠癌患者预后的关系,采用GO和KEGG分析ATP2A1表达水平可能参与的细胞功能及信号通路。ESTIMATE算法分析ATP2A1表达对肿瘤微环境及肿瘤突变负荷的影响。CCK8增殖实验及流式细胞周期实验检测ATP2A1参与结直肠癌增殖的机制,采用划痕伤口愈合实验、Transwell实验和Western blot实验分析ATP2A1参与结直肠癌迁移的机制。结果:依据ATP2A1表达的中位值将结直肠癌患者分为高、低表达两组,Kaplan-Meier曲线及Cox回归分析表明ATP2A1高表达是结直肠癌患者的预后风险因素,ESTIMATE算法结果表明ATP2A1表达降低免疫细胞浸润丰度,增加肿瘤突变负荷(TMB)和肿瘤细胞干性。CCK8增殖实验及细胞周期检测实验结果表明敲低ATP2A1表达引起直肠癌细胞S期阻滞进而抑制细胞增殖。划痕伤口愈合实验、Transwell和Western blot实验结果表明ATP2A1可以通过促进结直肠癌细胞上皮间充质转化增强结直肠癌细胞的迁移能力。结论:ATP2A1在结直肠癌中高表达,促进结直肠癌细胞的增殖及迁移,影响结直肠癌患者的预后和免疫治疗效果。

Objective:To investigate the mechanism of endoplasmic reticulum Ca2+-translocating ATPase isoform 1(ATP2A1)involved in influencing the biological behavior and immune infiltration of colorectal cancer(CRC).Methods:Transcriptome and immunohistochemistry(IHC)were used to analyze the relationship between ATP2A1 expression in colorectal cancer tumors and the prognosis of colorectal cancer patients,and GO and KEGG were used to analyze the cellular functions and signaling pathways that may be involved in the expression level of ATP2A1.The ESTIMATE algorithm was used to analyze the effect of ATP2A1 expression on the tumor microenvironment and the mutational load of tumors.The CCK8 proliferation assay and flow cell cycle assay were used to detect the mechanism of ATP2A1 involved in colorectal cancer proliferation,and the mechanism of ATP2A1 involved in colorectal cancer migration was analyzed by using scratch wound healing assay,Transwell assay and Western blot assay.Results:Colorectal cancer patients were divided into two groups of high and low expression based on the median value of ATP2A1 expression.Kaplan-Meier curves and Cox regression analyses demonstrated that high ATP2A1 expression was a prognostic risk factor for colorectal cancer patients,and the results of the ESTIMATE algorithm demonstrated that ATP2A1 expression decreased the abundance of immune cell infiltration,increased the tumor mutational load(TMB)and tumor cell stemness.The results of CCK8 proliferation assay and cell cycle assay showed that knockdown of ATP2A1 expression caused S-phase blockage in rectal cancer cells and thus inhibited cell proliferation.The results of scratch wound healing assay,Transwell and Western blot assay showed that ATP2A1 could enhance the migration ability of colorectal cancer cells by promoting epithelial mesenchymal transition of colorectal cancer cells.Conclusion:ATP2A1 is highly expressed in colorectal cancer,which can promote the proliferation and migration of colorectal cancer cells,and affect the prognosis and immunotherapeutic efficacy of colorectal cancer patients.