Table10_Single-Cell RNA Sequencing Reveals the Role of Phosphorylation-Related Genes in Hepatocellular Carcinoma Stem Cells.XLSX

Abnormal activation of protein kinases and phosphatases is implicated in various tumorigenesis, including hepatocellular carcinoma (HCC). Advanced HCC patients are treated with systemic therapy, including tyrosine kinase inhibitors, which extend overall survival. Investigation of the underlying mechanism of protein kinase signaling will help to improve the efficacy of HCC therapy. Combining single-cell RNA sequencing data and TCGA RNA-seq data, we profiled the protein kinases, phosphatases, and other phosphorylation-related genes (PRGs) of HCC patients in this study. We found nine protein kinases and PRGs with high expression levels that were mainly detected in HCC cancer stem cells, including POLR2G, PPP2R1A, POLR2L, PRC1, ITBG1BP1, MARCKSL1, EZH2, DTYMK, and AURKA. Survival analysis with the TCGA dataset showed that these genes were associated with poor prognosis of HCC patients. Further correlation analysis showed that these genes were involved in cell cycle-related pathways that may contribute to the development of HCC. Among them, AURKA and EZH2 were identified as two hub genes by Ingenuity Pathway Analysis. Treatment with an AURKA inhibitor (alisertib) and an EZH2 inhibitor (gambogenic) inhibited HCC cell proliferation, migration, and invasion. We also found that both AURKA and EZH2 were highly expressed in TP53-mutant HCC samples. Our comprehensive analysis of PRGs contributes to illustrating the mechanisms underlying HCC progression and identifying potential therapeutic targets for future clinical trials.

蛋白激酶与磷酸酶的异常激活与多种肿瘤发生密切相关，包括肝细胞癌（hepatocellular carcinoma, HCC）。晚期肝细胞癌患者的系统性治疗方案包含酪氨酸激酶抑制剂，该类药物可延长患者总生存期。深入解析蛋白激酶信号通路的潜在机制，有助于提升肝细胞癌治疗的临床疗效。本研究结合单细胞RNA测序（single-cell RNA sequencing）数据与癌症基因组图谱（The Cancer Genome Atlas, TCGA）RNA测序数据，对肝细胞癌患者的蛋白激酶、磷酸酶及其他磷酸化相关基因（phosphorylation-related genes, PRGs）进行了表达谱分析。我们筛选出9个在肝细胞癌肿瘤干细胞中高表达的蛋白激酶及磷酸化相关基因，分别为POLR2G、PPP2R1A、POLR2L、PRC1、ITBG1BP1、MARCKSL1、EZH2、DTYMK及AURKA。基于TCGA数据集的生存分析结果显示，上述基因与肝细胞癌患者的不良预后显著相关。进一步的相关性分析表明，这些基因参与细胞周期相关通路，可能在肝细胞癌的发生发展中发挥驱动作用。其中，通过Ingenuity通路分析（Ingenuity Pathway Analysis, IPA），我们鉴定出AURKA与EZH2为两个核心枢纽基因。分别使用AURKA抑制剂阿利塞利布（alisertib）与EZH2抑制剂藤黄酸（gambogenic）处理肝细胞癌细胞，可抑制其增殖、迁移及侵袭能力。我们还发现，在TP53突变型肝细胞癌样本中，AURKA与EZH2均呈高表达状态。本研究对磷酸化相关基因的全面分析，有助于阐明肝细胞癌进展的潜在分子机制，并为未来临床试验筛选潜在的治疗靶点。