Yu S H, Lee C M, Ha S H, Lee J, Jang K Y, Park S H
Department of Bio and Chemical Engineering, 65686Hongik University, Sejong, Republic of Korea.
Division of Biotechnology, 26714Jeonbuk National University, Iksan, Republic of Korea.
Hum Exp Toxicol. 2021 Feb;40(2):231-244. doi: 10.1177/0960327120943935. Epub 2020 Aug 13.
Tomentosin, a sesquiterpene lactone, is known to possess various biological activities. However, its anticarcinogenic activity against human hepatocellular carcinoma (HCC) cells has not been investigated in detail. Thus, this study aimed to elucidate the cytotoxic mechanism of tomentosin in human HCC cell lines HepG2 and Huh7. WST-1, cell counting, and colony formation assay results showed that treatment with tomentosin decreased the viability and suppressed the proliferation rate of HepG2 and Huh7 cells in a dose- and time-dependent manner. Cell cycle analysis revealed increased population of cells at the SubG1 and G2/M stage, and decreased population of cells at the G0/1 stage in HepG2 and Huh7 cells treated with tomentosin. Annexin V/propidium iodide double staining and TUNEL assay results showed increased apoptotic cell population and DNA fragmentation in HepG2 and Huh7 cells treated with tomentosin. Western blotting analysis results showed that tomentosin treatment significantly increased the expression level of Bax, Bim (short form), cleaved PARP1, FOXO3, p53, pSer15p53, pSer20p53, pSer46p53, p21, and p27, but decreased the expression of Bcl2, caspase3, caspase7, caspase9, cyclin-dependent kinase 2 (CDK2), CDK4, CDK6, cyclinB1, cyclinD1, cyclinD2, cyclinD3, and cyclinE in a dose-dependent manner. Taken together, this study revealed that tomentosin, which acted through cell cycle arrest and apoptosis, may be a useful therapeutic option against HCC.
托美辛是一种倍半萜内酯，已知具有多种生物活性。然而，其对人肝癌（HCC）细胞的抗癌活性尚未得到详细研究。因此，本研究旨在阐明托美辛在人肝癌细胞系HepG2和Huh7中的细胞毒性机制。WST-1、细胞计数和集落形成试验结果表明，托美辛处理以剂量和时间依赖性方式降低了HepG2和Huh7细胞的活力并抑制了其增殖率。细胞周期分析显示，在用托美辛处理的HepG2和Huh7细胞中，SubG1和G2/M期细胞数量增加，G0/1期细胞数量减少。膜联蛋白V/碘化丙啶双染色和TUNEL试验结果表明，在用托美辛处理的HepG2和Huh7细胞中，凋亡细胞数量增加且DNA片段化。蛋白质印迹分析结果表明，托美辛处理显著增加了Bax、Bim（短形式）、裂解的PARP1、FOXO3、p53、pSer15p53、pSer20p53、pSer46p53、p21和p27的表达水平，但以剂量依赖性方式降低了Bcl2、caspase3、caspase7、caspase9、细胞周期蛋白依赖性激酶2（CDK2）、CDK4、CDK6、细胞周期蛋白B1、细胞周期蛋白D1、细胞周期蛋白D2、细胞周期蛋白D3和细胞周期蛋白E的表达。综上所述，本研究表明，通过细胞周期阻滞和凋亡发挥作用的托美辛可能是一种治疗肝癌的有效选择。
