Abstract:

Objective　To investigate the mechanism by which ethyl acetate extract from Jiedu Xiaozheng Yin (EE-JXY) inhibits the growth of hepatocellular carcinoma cells, specifically focusing on side population (SP) cells in hepatocellular carcinoma. Methods　This study was from January to May 2023. Human Huh7 and PLC/PRF/5 cell lines were cultured in vitro. The presence of SP cells in the Huh7 and PLC/PRF/5 cell lines was determined and subsequently isolated by the flow cytometry. The colony formation ability of the SP cells was detected by the plate cloning assay. The SP cells' ability to form spheres in vitro was evaluated by the clonal sphere-forming assay. The effect of EE-JXY on the β-catenin protein translocation in the SP cells of hepatocellular carcinoma was detected by immunofluorescence. Results　The SP cells' quantity in the EE-JXY group was significantly lower than that in the blank group. The EE-JXY group showed a significant inhibition in the clonal formation and pellet formation abilities of the SP cells in vitro. The results of immunofluorescence showed that the β-catenin in the cytoplasm and nucleus was translocated to the cell membrane in some SP cells by EE-JXY. Conclusions　EE-JXY can significantly inhibit the proliferation of SP cells in Huh7 and PLC/PRF/5 cell lines; the mechanism may be related to the inhibition of β-catenin translocation.

Key words:

Hepatocellular carcinoma, Jiedu Xiaozheng Yin, Side population cells, β-catenin

摘要：

目的　从肝癌侧群（side population，SP）细胞的角度探讨解毒消癥饮（Jiedu Xiaozheng Yin，JXY）乙酸乙酯提取物（ethyl acetate extract from JXY，EE-JXY）抑制肝癌细胞生长的作用机制。方法　研究时间2023年1月至5月。体外培养人肝癌细胞株Huh7和PLC/PRF/5，流式细胞仪分析肝癌细胞中SP细胞的含量并分选SP细胞；平板克隆实验检测SP细胞的克隆形成能力；克隆球形成实验检测SP细胞的体外成球能力；免疫细胞荧光染色技术检测EE-JXY对肝癌SP细胞β-catenin蛋白转位的影响。结果　EE-JXY组SP细胞的含量显著低于空白组；SP细胞克隆形成能力和体外成球能力均受到显著抑制；EE-JXY使部分SP细胞β-catenin的表达从胞浆和细胞核转位到了细胞膜。结论　EE-JXY能显著抑制Huh7和PLC/PRF/5肝癌SP细胞的增殖，其机制可能与其抑制β-catenin的转位有关。

关键词:

肝癌, 解毒消癥饮, 侧群细胞, β-catenin