Coptisine, extracted from rhizoma coptidis, has been shown to inhibit a variety of cancers. However, the underlying mechanism by which coptisine regulates hepatocellular carcinoma (HCC) progression remains unknown. MTT assay, transwell invasion assay, and TUNEL assay were employed to examine cell viability, invasion, and apoptosis. In vivo tumor growth was determined by xenograft experiment. Reverse transcription-quantitative PCR was used to detect circCCT3 and HK2 gene expression. We utilized glucose consumption and lactate production assay to examine glucose metabolism state. Conditioned medium of coptisine-treated cancer-associated fibroblast (CAF) suppressed cell viability and invasion of HepG2 and Huh-7, whereas increased cell apoptosis. Coptisine significantly inhibited tumor growth of HepG2 cells in immunodeficient mice. Mechanistically, coptisine blocked secretion of exosomal circCCT3 from CAF. More notably, circCCT3 was upregulated in clinical HCC tumors. Moreover, circCCT3 was confirmed to affect glucose metabolism of HCC cells. We identified HK2 as a key downstream effector for circCCT3-modulated HCC tumorigenesis. In summary, our research revealed novel molecular mechanism of coptisine-blocked HCC progression, thereby providing solid rationale for using coptisine to treat HCC.

中文翻译：

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黄连碱阻断癌症相关成纤维细胞分泌外泌体 circCCT3 以重新编程肝细胞癌中的葡萄糖代谢


黄连碱是从黄连中提取的，已被证明可以抑制多种癌症。然而，黄连碱调节肝细胞癌（HCC）进展的潜在机制仍不清楚。采用MTT法、Transwell侵袭法和TUNEL法检测细胞活力、侵袭和凋亡。通过异种移植实验测定体内肿瘤生长。采用逆转录定量PCR检测circCCT3和HK2基因表达。我们利用葡萄糖消耗和乳酸产生测定来检查葡萄糖代谢状态。黄连碱处理的癌症相关成纤维细胞 (CAF) 的条件培养基抑制了 HepG2 和 Huh-7 的细胞活力和侵袭，同时增加了细胞凋亡。黄连碱显着抑制免疫缺陷小鼠 HepG2 细胞的肿瘤生长。从机制上讲，黄连碱可阻断 CAF 分泌外泌体 circCCT3。更值得注意的是，circCCT3 在临床 HCC 肿瘤中表达上调。此外，circCCT3被证实影响HCC细胞的葡萄糖代谢。我们确定 HK2 是 circCCT3 调节的 HCC 肿瘤发生的关键下游效应子。总之，我们的研究揭示了黄连碱阻断 HCC 进展的新分子机制，从而为使用黄连碱治疗 HCC 提供了坚实的理论基础。