Potential Anticancer Activity of the Furanocoumarin Derivative Xanthotoxin Isolated from Ammi majus L. Fruits: In Vitro and In Silico Studies.

Marwa Y Issa,Mohamed F Elshal,Mokhtar Bishr,Yasmin S Abulfadl,Osama Salama,Noha Fathallah,Mostafa A Abdelkawy

doi:10.3390/molecules27030943

Marwa Y Issa, Mohamed F Elshal + Show 5 more

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https://doi.org/10.3390/molecules27030943

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Abstract

Ammi majus 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 Ammi majus fruit methanol extract (AME) against liver cancer and to elucidate the active compound(s) and their mechanisms of action. Three fractions from AME (Hexane, CH2Cl2, 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, CH2Cl2 fraction revealed the most potent cytotoxic activity. The structures of compounds isolated from the CH2Cl2 fraction were elucidated using 1H- and 13C-NMR and found that Compound 1 (xanthotoxin) has the strongest cytotoxic activity against HepG2 cells (IC50 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.

Highlights

Hepatocellular carcinoma (HCC) is the sixth and the fourth common cancer in the world and in Egypt, respectively [1]
These results suggest that xanthotoxin negatively affected autophagy
The sulforhodamine B (SRB) assay was applied to determine the antiproliferative activity of the different isolated compounds, and it was found that the coumarin xanthotoxin has the highest cytotoxic activity, of almost three-fold and one-fold greater compared to total Ammi majus fruit methanol extract (AME) and CH2Cl2 fractions, respectively, approaching that of doxorubicin with IC50 6.9 μg/mL

Summary

Introduction

Hepatocellular carcinoma (HCC) is the sixth and the fourth common cancer in the world and in Egypt, respectively [1]. HCC represents one of the most aggressive solid tumors and the third leading cause of cancer-related mortality worldwide. The prognosis of patients with HCC is poor, with around 80% mortality rate within one year from the first diagnosis and an overall 5-years survival rate of 18% [2]. Despite decades of basic and clinical research and trials of systemic chemotherapy for HCC including cisplatin, doxorubicin, and 5-flurouracil, which are used as mono- or combined therapy, only 10 to 25% response rate with marginal survival improvement have been reached mainly due to development of resistance and recurrence [3]. Among the other advantages of natural drugs are their easy availability and economic value; the climbing demands for these rich and potent agents is based on their efficacy to scavenge free radicals, stimulate apoptosis, and inhibit cancer cells’ survival [7]

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