Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt.
Molecular Biology Department, Genetic Engineering and Biotechnology Institute, University of Sadat City, Sadat City 32897, Egypt.
Molecules. 2022 Jan 29;27(3):943. doi: 10.3390/molecules27030943.
L., an indigenous plant in Egypt, is widely used in traditional medicine due to its various pharmacological properties. We aimed to evaluate the anticancer properties of fruit methanol extract (AME) against liver cancer and to elucidate the active compound(s) and their mechanisms of action. Three fractions from AME (Hexane, CHCl, and EtOAc) were tested for their anticancer activities against HepG2 cell line in vitro (cytotoxicity assay, cell cycle analysis, annexin V-FITC apoptosis assay, and autophagy efflux assay) and in silico (molecular docking). Among the AME fractions, CHCl fraction revealed the most potent cytotoxic activity. The structures of compounds isolated from the CHCl fraction were elucidated using H- and C-NMR and found that Compound (xanthotoxin) has the strongest cytotoxic activity against HepG2 cells (IC 6.9 ± 1.07 µg/mL). Treating HepG2 cells with 6.9 µg/mL of xanthotoxin induced significant changes in the DNA-cell cycle (increases in apoptotic pre-G1 and G2/M phases and a decrease in the S-phase). Xanthotoxin induced significant increase in Annexin-V-positive HepG2 cells both at the early and late stages of apoptosis, as well as a significant decrease in autophagic flux in cancer compared with control cells. In silico analysis of xanthotoxin against the DNA-relaxing enzyme topoisomease II (PDB code: 3QX3) revealed strong interaction with the key amino acid Asp479 in a similar fashion to that of the co-crystallized inhibitor (etoposide), implying that xanthotoxin has a potential of a broad-spectrum anticancer activity. Our results indicate that xanthotoxin exhibits anticancer effects with good biocompatibility toward normal human cells. Further studies are needed to optimize its antitumor efficacy, toxicity, solubility, and pharmacokinetics.
L. 是埃及的一种本土植物，由于其多种药理学特性，在传统医学中被广泛应用。我们旨在评估 果实甲醇提取物（AME）对肝癌的抗癌特性，并阐明其活性化合物及其作用机制。我们测试了 AME 的三种馏分（己烷、CHCl 和 EtOAc）对 HepG2 细胞系的体外抗癌活性（细胞毒性测定、细胞周期分析、Annexin V-FITC 凋亡测定和自噬流出测定）和计算机模拟（分子对接）。在 AME 馏分中，CHCl 馏分显示出最强的细胞毒性活性。从 CHCl 馏分中分离得到的化合物的结构通过 H-NMR 和 C-NMR 阐明，发现化合物 （花椒毒素）对 HepG2 细胞具有最强的细胞毒性（IC 6.9 ± 1.07 µg/mL）。用 6.9 µg/mL 的花椒毒素处理 HepG2 细胞，导致 DNA 细胞周期发生显著变化（凋亡前 G1 和 G2/M 期增加，S 期减少）。花椒毒素诱导 Annexin-V 阳性 HepG2 细胞凋亡早期和晚期的显著增加，以及与对照细胞相比，癌细胞中自噬通量的显著降低。花椒毒素与松弛酶拓扑异构酶 II（PDB 代码：3QX3）的计算机模拟分析显示，其与关键氨基酸 Asp479 的强烈相互作用与共晶抑制剂（依托泊苷）相似，这表明花椒毒素具有广谱抗癌活性的潜力。我们的结果表明，花椒毒素表现出抗癌作用，对正常人类细胞具有良好的生物相容性。需要进一步研究来优化其抗肿瘤疗效、毒性、溶解度和药代动力学。
