YWHAB基因通过PI3K/AKT通路调控肝细胞癌增殖和侵袭并重塑免疫抑制微环境

YWHAB gene regulates hepatocellular carcinoma proliferation and invasion via the PI3K/AKT pathway and remodels the immunosuppressive microenvironment

目的:探究PANoptosis关键基因YWHAB在肝细胞癌(hepatocellular carcinoma,HCC)中的表达水平和作用机制及其与肿瘤免疫微环境的关系。方法:来自南通大学附属医院肝胆胰脾外科的组织芯片和癌症基因组图谱(the Cancer Genome Atlas,TCGA)数据库分析YWHAB在HCC中的表达及与预后的关系。体外构建HCC的PLC/PRF/5和Huh7的YWHAB敲低以及过表达模型,评估其肿瘤细胞的增殖和侵袭能力,并用Western Blot验证其通路。通过ssGSEA/TIDE/ESTIMATE 3种免疫浸润算法分析YWHAB对免疫环境的影响,并用体外CD8+T细胞杀伤实验进行验证。结果:YWHAB在HCC组织中阳性表达率为70.46%,在癌旁组织阳性表达率为20.46%,差异有统计学意义(P<0.05),YWHAB高表达与肿瘤包膜完整性、血管浸润、淋巴结转移、肿瘤分化程度和晚期TNM分期均明显相关(均P<0.05)。Kaplan-Meier生存分析表明,YWHAB高表达患者总体生存(overall survival,OS)率明显降低(P=0.003)。体外实验证明,YWHAB能通过PI3K/AKT信号通路调控肿瘤细胞的增殖和侵袭。免疫微环境方面,YWHAB表达与辅助型T细胞2(R=0.402,P<0.05)、巨噬细胞(R=0.321,P<0.05)等抑制性免疫细胞浸润数量呈正相关,与免疫耗竭性检查标志物CD274、HAVCR2、PDCD1、TIGIT、LAG3和CTLA4等表达呈正相关。YWHAB低表达组显示出更好的免疫治疗敏感评分,当敲低YWHAB基因后,T细胞对肿瘤细胞的杀伤效果显著增强(P<0.05)。结论:YWHAB作为HCC的癌基因和不良预后标志物,可能通过调控PI3K/AKT通路途径而促进HCC的增殖和侵袭,且与免疫抑制微环境相关。

Objective:To explore the expression levels and functional mechanisms of YWHAB,a key gene in PANoptosis,in hepatocellular carcinoma(HCC),and its relationship with the tumor immune microenvironment.Methods:Tissue microarrays from the Department of Hepatobiliary and Pancreatic Surgery of the Affiliated Hospital of Nantong University and the Cancer Genome Atlas(TCGA)database were used to analyze the expression and prognostic association of YWHAB in HCC.In vitro models of YWHAB knockdown and overexpression in HCC PLC/PRF/5 and Huh7 cells were established to evaluate its influence on tumor cell proliferation and invasion,with its pathway validated by Western Blot.The impact of YWHAB on the immune environment was analyzed using three immune infiltration algorithms:ssGSEA/TIDE/ESTIMATE,and further validated by in vitro CD8+T cell cytotoxicity assay.Results:The positive expression rate of YWHAB in HCC tissues was 70.46%,significantly higher than the 20.46%in adjacent tissues(P<0.05).High expression of YWHAB was significantly associated with tumor capsule integrity,vascular invasion,lymph node metastasis,tumor differentiation,and advanced TNM stage(P<0.05 for all).Kaplan-Meier survival analysis revealed that high YWHAB expression was associated with significantly reduced overall survival(OS)rate(P=0.003).In vitro experiments demonstrated that YWHAB regulates tumor cell proliferation and invasion via the PI3K/AKT signaling pathway.Regarding the immune landscape,YWHAB expression was positively correlated with the infiltration of immunosuppressive cells such as T helper 2 cells(R=0.402,P<0.05)and macrophages(R=0.321,P<0.05),and the expression of immune exhaustion markers including CD274,HAVCR2,PDCD1,TIGIT,LAG3,and CTLA4.The low YWHAB expression group showed a higher immune treatment sensitivity score,and the cytotoxicity of T cells to tumor cells significantly increased after YWHAB knockdown(P<0.05).Conclusion:YWHAB acts as an oncogene and adverse prognosis indicator in HCC,possibly promoting HCC proliferation and invasion by regulating the PI3K/AKT pathway,and is associated with an immunosuppressive microenvironment.