6,8-Diprenylorobol is a phytochemical derived from the roots of Glycyrrhiza uralensis Fisch. 6,8-Diprenylorobol exhibits several biological activities, but the effects of 6,8-diprenylorobol on cancers have been hardly investigated. This study is aimed at elucidating the anticancer effect and working mechanism of 6,8-diprenylorobol in HepG2 and Huh-7, two kinds of human hepatocellular carcinoma (HCC) cell lines. WST-1, cell counting, and colony formation assays and morphological change analysis showed that 6,8-diprenylorobol treatment decreased the cell viability and proliferation rate. Cell cycle analysis indicated that 6,8-diprenylorobol treatment increased the population of the G1/0 stage. Annexin V/PI double staining and TUNEL analysis showed that 6,8-diprenylorobol treatment increased the apoptotic cell population and DNA fragmentation. Western blot analysis showed that 6,8-diprenylorobol treatment increased the expression of cleaved PARP1, cleaved caspase-3, FOXO3, Bax, Bim, p21, and p27 but decreased the expression of Bcl2 and BclXL. Interestingly, 6,8-diprenylorobol inhibited CYP2J2-mediated astemizole O-demethylation and ebastine hydroxylase activities with values of 9.46 and 2.61 μM, respectively. CYP2J2 siRNA transfection enhanced the anticancer effect of 6,8-diprenylorobol in HepG2 and Huh-7 cells through the downregulation of CYP2J2 protein expression and upregulation of FOXO3. Taken together, this study proposes that 6,8-diprenylorobol treatment may be a useful therapeutic option against HCC by targeting CYP2J2 and FOXO3.

中文翻译：

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6,8-Diprenylorobol 通过激活 FOXO3 和抑制 CYP2J2 诱导人肝细胞癌细胞凋亡

6,8-Diprenylorobol 是一种来自甘草根的植物化学物质菲施。6,8-Diprenylorobol 表现出多种生物活性，但几乎没有研究过 6,8-diprenylorobol 对癌症的影响。本研究旨在阐明6,8-diprenylorobol在两种人肝细胞癌（HCC）细胞系HepG2和Huh-7中的抗癌作用和作用机制。WST-1、细胞计数、集落形成试验和形态变化分析表明，6,8-diprenylorobol 处理降低了细胞活力和增殖率。细胞周期分析表明 6,8-diprenylorobol 治疗增加了 G1/0 阶段的人口。Annexin V/PI 双染色和 TUNEL 分析表明 6,8-diprenylorobol 处理增加了凋亡细胞群和 DNA 片段化。蛋白质印迹分析表明，6, 8-diprenylorobol 治疗增加了裂解的 PARP1、裂解的 caspase-3、FOXO3、Bax、Bim、p21 和 p27 的表达，但降低了 Bcl2 和 BclXL 的表达。有趣的是，6,8-diprenylorobol 抑制 CYP2J2 介导的阿司咪唑Ò -demethylation和依巴斯汀与羟化酶活性的9.46和2.61的值 μ分别男，。CYP2J2 siRNA 转染通过下调 CYP2J2 蛋白表达和上调 FOXO3 来增强 6,8-diprenylorobol 在 HepG2 和 Huh-7 细胞中的抗癌作用。综上所述，该研究表明，通过靶向 CYP2J2 和 FOXO3，6,8-diprenylorobol 治疗可能是一种有效的 HCC 治疗选择。