Abdul-Nabie Hoda, Samir Safia, Aboshousha Tarek, Fares Nagui, Abo-Zeid Faten Sabra
Zoology Department, Faculty of Science, Ain Shams University, Egypt.
Biochemistry and Molecular Biology Department, Theodor Bilharz Research Institute, Giza, Egypt.
Toxicol Rep. 2025 Jul 19;15:102091. doi: 10.1016/j.toxrep.2025.102091. eCollection 2025 Dec.
Hepatocellular carcinoma (HCC) is a major public health problem, with a poor prognosis in patients with advanced disease. We aimed to investigate the cytotoxic activity of coumestrol against human hepatocellular carcinoma HepG2 cells. Crystal violet (CV) assay was performed for cell cytotoxicity assessment on Vero cells (normal Kidney) and HepG2 cells. Apoptosis was analyzed by Annexin V/FITC staining. The relative expression of BAX, bcl-2, NF-Kβ, PCNA, MMP2, Caspase-3, Caspase-9, COX2, and MMP9 was detected using quantitative real-time PCR (qPCR). In addition, immunohistochemical analysis (IHC) of Bax, and PCNA were established. Results indicated that coumestrol induced significant toxicity in HepG2 cells. Annexin V-FITC staining assays revealed that coumestrol-induced cytotoxicity in HepG2 cells was mediated through apoptosis stimulation. The apoptosis in HepG2 cells was mediated through caspase-activation. Cell invasion was inhibited by coumestrol in HepG2 cells via inhibition of MMP-2 and MMP-9 expressions. IHC confirmed the strong expression of Bax and nuclear expression of PCNA in treated cells. To the best of our knowledge, limited studies have investigated the impact of coumestrol. The study showed that coumestrol exhibited cytotoxic and apoptotic effects against HepG2 cells, accompanied by inhibition of invasion-related gene expression.
肝细胞癌（HCC）是一个重大的公共卫生问题，晚期疾病患者的预后较差。我们旨在研究香豆雌酚对人肝细胞癌HepG2细胞的细胞毒性活性。采用结晶紫（CV）试验对Vero细胞（正常肾细胞）和HepG2细胞进行细胞毒性评估。通过膜联蛋白V/异硫氰酸荧光素（FITC）染色分析细胞凋亡情况。使用定量实时聚合酶链反应（qPCR）检测BAX、bcl-2、核因子κB（NF-κβ）、增殖细胞核抗原（PCNA）、基质金属蛋白酶2（MMP2）、半胱天冬酶-3（Caspase-3）、半胱天冬酶-9（Caspase-9）、环氧化酶2（COX2）和基质金属蛋白酶9（MMP9）的相对表达。此外，还进行了Bax和PCNA的免疫组织化学分析（IHC）。结果表明，香豆雌酚对HepG2细胞具有显著毒性。膜联蛋白V-FITC染色试验显示，香豆雌酚诱导HepG2细胞的细胞毒性是通过刺激细胞凋亡介导的。HepG2细胞的凋亡是通过半胱天冬酶激活介导的。香豆雌酚通过抑制MMP-2和MMP-9的表达抑制HepG2细胞的侵袭。免疫组织化学分析证实，处理后的细胞中Bax表达强烈，PCNA呈核表达。据我们所知，对香豆雌酚影响的研究有限。该研究表明，香豆雌酚对HepG2细胞具有细胞毒性和凋亡作用，并伴有对侵袭相关基因表达的抑制。
