我们分析了在肝细胞癌（HCC）中TMEM64的表达，并研究了TMEM64表达水平对HCC细胞增殖和侵袭的影响。我们根据TCGA和GTEx数据库的数据分析了HCC和相邻组织中TMEM64的表达水平。使用Kaplan-Meier生存分析和Cox回归模型来检验TMEM64对HCC患者的预后价值，并基于TMEM64的表达和患者的临床特征构建了一个Nomogram。进行功能富集分析以探索潜在的信号通路，并使用单样本基因集富集分析评估免疫细胞浸润情况。我们还进行了细胞实验，使用CCK-8、EdU、集落形成、Transwell和愈合缝合实验观察了HCCLM3细胞中TMEM64沉默以及Huh7细胞中TMEM64过表达后增殖、迁移和侵袭的变化。 TMEM64的表达水平在HCC中显著高于相邻组织（P < 0.05）。Kaplan-Meier分析表明，TMEM64高表达与患者不良预后相关（P < 0.05）。多元Cox回归分析表明，高TMEM64表达是HCC患者总体生存的独立风险因素（P < 0.05）。TMEM64的表达水平与NK细胞、CD8+ T细胞和浆细胞型pDCs细胞的免疫细胞浸润水平呈负相关（P < 0.05）。GO、KEGG和GSEA富集分析显示TMEM64在肿瘤侵袭和转移等通路中显著富集。Nomogram和校准曲线显示该模型的预测可靠性中等。在细胞实验中，TMEM64沉默明显抑制，而TMEM64过表达显著促进了HCC细胞的增殖、迁移和侵袭（P < 0.01）。 高TMEM64表达可能作为HCC不良预后的独立风险因素，并促进HCC细胞在体外的增殖、迁移和侵袭。

To analyze the expression of TMEM64 in hepatocellular carcinoma (HCC) and investigate the effect of TMEM64 expression level on proliferation and invasion of HCC cells in vitro.We analyzed the expression level of TMEM64 in HCC and adjacent tissues based on data from TCGA and GTEx databases. The prognostic value of TMEM64 for HCC patients was examined using Kaplan-Meier survival analysis and a Cox regression model, and a nomogram was constructed based on TMEM64 expression and clinical characteristics of the patients. Functional enrichment analysis was performed to explore the potential signaling pathways, and immune cell infiltration was assessed using single sample gene set enrichment analysis. We also performed cell experiment to observe the changes in proliferation, migration, and invasion in HCCLM3 cells with TMEM64 knockdown and in Huh7 cells with TMEM64 overexpression using CCK-8, EdU, colony formation, Transwell, and wound healing assays.The expression level of TMEM64 was significantly higher in HCC than in the adjacent tissues (P < 0.05). Kaplan-Meier analysis suggested that a high expression of TMEM64 was associated with poor outcomes of the patients (P < 0.05). Multivariate Cox regression analysis indicated that a high TMEM64 expression was an independent risk factor for overall survival of HCC patients (P < 0.05). TMEM64 expression level was negatively correlated with the levels of immune cell infiltration by NK cells, CD8 + T cells, and plasma pDCs cells (P < 0.05). GO, KEGG, and GSEA enrichment analyses showed that TMEM64 was significantly enriched with tumor invasion and metastasis pathways. The nomogram and calibration curves indicated a moderate prediction reliability of the model. In the cell experiment, TMEM64 knockdown obviously suppressed and TMEM64 overexpression markedly promoted the proliferation, migration, and invasion of HCC cells (P < 0.01).A high TMEM64 expression may serve as an independent risk factor for poor prognosis of HCC and promotes proliferation, migration, and invasion of HCC cells in vitro.