Extended Data Fig. 4: Pseudohypoxia drives the metastatic potential of THY1+ CSCs.
a, Schematic overview of datasets used in pan-cancer scRNA analysis. b–d, Metastasis score (b) and hypoxia signature score (c) of THY1+ CSCs in pan-cancer scRNA datasets. The correlation of average metastasis score and hypoxia signature score across different cancer types (d). e, Distribution of hypoxic area (stained by hypoxyprobe) and HIF1α+ cells in tumours of Thy1-OE Hepa1-6 hepatoma-bearing mice (n = 11; scale bar, 75 µm). The numbers of HIF1α+ cells in normoxic and hypoxic areas were calculated. f,g, Venn diagram showing genes related to pseudohypoxia in Hepa1-6 hepatoma, defined as those upregulated in Thy1-OE versus WT mouse hepatoma, as well as in Hif1a-competent versus Hif1a-knockdown Thy1-OE groups (f). Cancer hallmark enrichment analysis of these genes was performed (g) using bulk RNA-seq data (n = 3; CRA036043). The red line indicates adjusted P < 0.05. h, Comparative analysis of metastasis-related gene expression profiles among THY1− CSCs, HIF-signaturehigh THY1+ CSCs, and HIF-signaturelow THY1+ CSCs in human HCC. i,j, Expression of hypoxia-related proteins (i) and EMT-related proteins (j) of WT and Thy1-OE Hepa1-6 cells was analysed (each n = 5). k, Migration potential of WT and Thy1-OE Hepa1-6 cells cultured in vitro was estimated (n = 5; scale bar, 60 µm). l, Effects of Thy1-OE on survival in mice bearing Hepa1-6 hepatomas (n = 8). Data are from three (f,g) or five (i,j) independent experiments. Data are presented as mean ± s.e.m. of three (e,l) or five (k) independent experiments. Statistical significance was determined using linear regression with two-sided t-tests for regression coefficients (d), over-representation analysis based on the hypergeometric test, with false discovery rate correction applied for multiple comparisons (g,h), unpaired two-tailed Student’s t-test (e,k) or the log-rank (Mantel–Cox) test (l). Violin plots show data distribution with median and interquartile range (b,c). Panel a created with BioRender.com. ERI, enabling replicative immortality; REM, reprogramming energy metabolism; RCD, resisting cell death; TPI, tumour-promoting inflammation; TIM, tissue invasion and metastasis; IA, inducing angiogenesis; AID, avoiding immune destruction; EGS, evading growth suppressors; GI, genome instability and mutation; SPS, sustaining proliferative signalling.
