Isocryptomerin (ISO) is a flavonoid isolated from the natural medicine Selaginellae Herba, which has various pharmacological activities. This study investigated the antitumor effect and underlying molecular mechanism of ISO on hepatocellular carcinoma (HCC) HepG2 cells. The cell viability assay revealed that ISO has a considerable killing effect on HCC cell lines. The apoptosis assay showed that ISO induced mitochondria‐dependent apoptosis through the Bad/cyto‐c/cleaved (cle)‐caspase‐3/cleaved (cle)‐PARP pathway. The network pharmacological analysis found 13 key target genes, and epidermal growth factor receptor (EGFR), AKT, mitogen‐activated protein kinase (MAPK), and reactive oxygen species (ROS) signaling pathways were strongly associated with ISO against HCC. Further verification of the results showed that ISO induced apoptosis by increasing p‐p38 and p‐JNK expression and decreasing p‐EGFR, p‐SRC, p‐ERK, and p‐STAT3 expression. Furthermore, ISO induced G0/G1 phase arrest by downregulating p‐AKT, Cyclin D, and CDK 4 expression and upregulating p21 and p27 expression in HepG2 cells. Moreover, ISO inhibited HepG2 cell migration by decreasing p‐GSK‐3β, β‐catenin, and N‐cadherin expression and increasing E‐cadherin expression. Additionally, ISO promoted ROS accumulation in HepG2 cells, and ISO‐induced apoptosis, arrest cell cycle, and inhibition of migration were reversed by an ROS scavenger, N‐acetyl‐ l‐cysteine. Overall, ISO induced cell apoptosis and cell cycle arrest and inhibited cell migration by ROS‐mediated EGFR, AKT, and MAPK signaling pathways in HepG2 cells.

中文翻译：

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异隐梅林对HepG2细胞抗肿瘤作用及分子机制的网络药理学分析及实验验证

异隐球菌素 (ISO) 是从天然药物中分离出来的黄酮类化合物卷柏亚科草本植物，具有多种药理活性。本研究探讨了 ISO 对肝细胞癌 (HCC) HepG2 细胞的抗肿瘤作用及其潜在分子机制。细胞活力测定表明ISO对HCC细胞系具有相当大的杀伤作用。细胞凋亡测定表明，ISO 通过 Bad/cyto-c/cleaved (cle)-caspase-3/cleaved (cle)-PARP 途径诱导线粒体依赖性细胞凋亡。网络药理学分析发现13个关键靶基因，表皮生长因子受体（EGFR）、AKT、丝裂原激活蛋白激酶（MAPK）和活性氧（ROS）信号通路与ISO抗HCC密切相关。进一步验证结果表明，ISO通过增加p-p38和p-JNK的表达，减少p-EGFR、p-SRC、p-ERK和p-STAT3的表达来诱导细胞凋亡。此外，ISO 通过下调 HepG2 细胞中 p-AKT、Cyclin D 和 CDK 4 的表达以及上调 p21 和 p27 的表达来诱导 G0/G1 期停滞。此外，ISO 通过减少 p-GSK-3β、β-catenin 和 N-cadherin 表达并增加 E-cadherin 表达来抑制 HepG2 细胞迁移。此外，ISO 促进了 HepG2 细胞中 ROS 的积累，并且 ISO 诱导的细胞凋亡、细胞周期停滞和迁移抑制被 ROS 清除剂 N-乙酰基-我‐半胱氨酸。总体而言，ISO 在 HepG2 细胞中诱导细胞凋亡和细胞周期停滞，并通过 ROS 介导的 EGFR、AKT 和 MAPK 信号通路抑制细胞迁移。