在癌症研究中，识别和发展导致肿瘤发展、复发、转移和耐药的癌症干细胞治疗靶点的重要目标。肝细胞癌细胞系Li-7含有功能不同的细胞类型。具有肿瘤形成活性的细胞在癌症干细胞样CD13+CD166-细胞中富集，而在常规培养基（含10%胎牛血清的RPMI1640）中，这个细胞群逐渐减少。当Li-7细胞在mTeSR1培养基中培养时，CD13+CD166-细胞及其肿瘤形成能力得以维持。在本研究中，我们试图鉴定该亚群体肿瘤形成能力的机制。我们比较了CD13+CD166-细胞与其他细胞亚群体的基因表达谱，并鉴定出CD13+CD166-细胞中过表达的九个基因（ENPP2, SCGN, FGFR4, MCOLN3, KCNJ16, SMIM22, SMIM24, SERPINH1, and TMPRSS2）。在从mTeSR1转移到含10%胎牛血清的RPMI1640培养基后，这九个基因在Li-7细胞中的表达减少，同时失去了肿瘤形成能力。相反，当这些基因在使用含10%胎牛血清的RPMI1640培养基进行强制表达的Li-7细胞培养中，Li-7细胞的肿瘤形成能力得以维持。使用毛细管电泳质谱分析的代谢组学分析显示"丙氨酸、天冬酰胺和谷氨酸代谢"和"精氨酸合成"两个代谢途径在癌症干细胞样细胞中被激活。本文的分析展示了治疗肝细胞癌癌症干细胞的潜在治疗靶点基因和代谢物。©2023. 作者。

The identification and development of therapeutic targets in cancer stem cells that lead to tumor development, recurrence, metastasis, and drug resistance is an important goal in cancer research. The hepatocellular carcinoma cell line Li-7 contains functionally different types of cells. Cells with tumor-forming activity are enriched in cancer stem cell-like CD13+CD166- cells and this cell population gradually decreases during culture in conventional culture medium (RPMI1640 containing 10% fetal bovine serum). When Li-7 cells are cultured in mTeSR1, a medium developed for human pluripotent stem cells, CD13+CD166- cells, and their tumorigenicity is maintained. Here, we sought to identify the mechanisms of tumorigenicity in this sub-population. We compared gene expression profiles of CD13+CD166- cells with other cell sub-populations and identified nine overexpressed genes (ENPP2, SCGN, FGFR4, MCOLN3, KCNJ16, SMIM22, SMIM24, SERPINH1, and TMPRSS2) in CD13+CD166- cells. After transfer from mTeSR1 to RPMI1640 containing 10% fetal bovine serum, the expression of these nine genes decreased in Li-7 cells and they lost tumorigenicity. In contrast, when these genes of Li-7 cells were forcibly expressed in cultures using RPMI1640 containing 10% fetal bovine serum, Li-7 cells maintained tumorigenicity. A metabolome analysis using capillary electrophoresis-mass spectrometry showed that two metabolic pathways, "Alanine, aspartate and glutamate metabolism" and "Arginine biosynthesis" were activated in cancer stem-cell-like cells. Our analyses here showed potential therapeutic target genes and metabolites for treatment of cancer stem cells in hepatocellular carcinoma.© 2023. The Author(s).