肿瘤异质性对理解肿瘤进展和转移的驱动机制提出了巨大的挑战。肝细胞癌（HCC）在细胞水平的异质性尚不清楚。对单细胞RNA测序数据和空间转录组数据进行整合分析。应用多种方法研究HCC肿瘤细胞的亚型。分析了不同亚型肿瘤细胞的功能特征、翻译因子、临床意义和微环境关联。分析亚型与成纤维细胞的相互作用。我们通过整合52个单细胞RNA测序数据和5个空间转录组数据建立了HCC恶性细胞的异质性景观。我们鉴定了肿瘤细胞的三种亚型，包括ARG1代谢亚型（Metab-亚型）、TOP2A增殖表型（Prol-表型）和S100A6促转移亚型（EMT-亚型）。富集分析发现，三种亚型具有不同的特征，即代谢、增殖和上皮间质转化。轨迹分析显示Metab亚型和EMT亚型均起源于Prol表型。翻译因子分析发现 EMT 亚型仅激活 SMAD3 和 TGF-β 信号通路。以EMT亚型细胞为主的HCC预后不良，微环境荒凉。我们发现肿瘤细胞和成纤维细胞之间存在由 SPP1-CD44 和 CCN2/TGF-β-TGFBR1 相互作用对介导的正循环。抑制 CCN2 会破坏循环，减轻向 EMT 亚型的转化，并抑制转移。通过建立恶性细胞的异质性景观，我们确定了 HCC 的三亚型分类。其中，S100A6肿瘤细胞在转移中发挥着至关重要的作用。针对肿瘤细胞和成纤维细胞之间的反馈环路是一种有前途的抗转移策略。© 2024。作者。

Tumor heterogeneity presents a formidable challenge in understanding the mechanisms driving tumor progression and metastasis. The heterogeneity of hepatocellular carcinoma (HCC) in cellular level is not clear.Integration analysis of single-cell RNA sequencing data and spatial transcriptomics data was performed. Multiple methods were applied to investigate the subtype of HCC tumor cells. The functional characteristics, translation factors, clinical implications and microenvironment associations of different subtypes of tumor cells were analyzed. The interaction of subtype and fibroblasts were analyzed.We established a heterogeneity landscape of HCC malignant cells by integrated 52 single-cell RNA sequencing data and 5 spatial transcriptomics data. We identified three subtypes in tumor cells, including ARG1+ metabolism subtype (Metab-subtype), TOP2A+ proliferation phenotype (Prol-phenotype), and S100A6+ pro-metastatic subtype (EMT-subtype). Enrichment analysis found that the three subtypes harbored different features, that is metabolism, proliferating, and epithelial-mesenchymal transition. Trajectory analysis revealed that both Metab-subtype and EMT-subtype originated from the Prol-phenotype. Translation factor analysis found that EMT-subtype showed exclusive activation of SMAD3 and TGF-β signaling pathway. HCC dominated by EMT-subtype cells harbored an unfavorable prognosis and a deserted microenvironment. We uncovered a positive loop between tumor cells and fibroblasts mediated by SPP1-CD44 and CCN2/TGF-β-TGFBR1 interaction pairs. Inhibiting CCN2 disrupted the loop, mitigated the transformation to EMT-subtype, and suppressed metastasis.By establishing a heterogeneity landscape of malignant cells, we identified a three-subtype classification in HCC. Among them, S100A6+ tumor cells play a crucial role in metastasis. Targeting the feedback loop between tumor cells and fibroblasts is a promising anti-metastatic strategy.© 2024. The Author(s).