Various populations of cancer stem cells (CSCs) have been identified in hepatocellular carcinoma (HCC). Wnt signaling is variably activated in HCC and regulates CSCs and tumorigenesis. We explored cell-to-cell Wnt and stemness heterogeneity in HCC by labeling freshly isolated cancer cells with a Wnt-specific reporter, thereby identifying a small subset (0.4%–8.9%) of Wnt-activity^high cells. Further cellular subset analysis identified a refined subset of Wnt-activity^highALDH1⁺EpCAM⁺ triple-positive (TP) cells as the most stem-like, phenotypically plastic, and tumorigenic among all putative CSC populations. These TP “superpotent CSCs” (spCSCs) specifically upregulate the expression of dishevelled 1 (DVL1) through the antagonism between abnormal spindle-like microcephaly-associated (ASPM) and the ubiquitin ligase complex Cullin-3/KLHL-12. Subsequent functional and molecular studies revealed the role of DVL1 in controlling spCSCs and their tumorigenic potential. These findings provide the mechanistic basis of the Wnt and stemness heterogeneity in HCC and highlight the important role of DVL1^high spCSCs in tumor progression.

在肝细胞癌（HCC）中已鉴定出多种癌症干细胞（CSC）群体。Wnt信号在HCC中被可变激活，并调节CSC和肿瘤发生。我们通过用Wnt特异性报告基因标记新鲜分离的癌细胞，探索了肝细胞中Wnt和干细胞异质性，从而鉴定了Wnt活性^高细胞的一小部分（0.4％–8.9％）。进一步的细胞亚群分析确定了Wnt活性^高的ALDH1 ⁺ EpCAM ⁺的精炼亚群三重阳性（TP）细胞是所有推定的CSC群体中最干的，表型可塑性且具有致瘤性的细胞。这些TP“超能CSC”（spCSC）通过异常纺锤样小头畸形相关（ASPM）与泛素连接酶复合体Cullin-3 / KLHL-12之间的拮抗作用，特异性上调了蓬乱的1（DVL1）的表达。随后的功能和分子研究揭示了DVL1在控制spCSCs中的作用及其致瘤潜力。这些发现提供了肝癌中Wnt和干异质性的机制基础，并突出了DVL1^高spCSCs在肿瘤进展中的重要作用。