Antiproliferative activity of ethyl acetate fraction of Euphorbia ingens against prostate cancer cell line: An in silico and in vitro analysis

Abstract

Current prostate cancer (PCa) treatments often lead to severe side effects, prompting the exploration of safer alternatives. Euphorbia ingens is a medicinal plant used for cancer treatment in African communities; however, there is no scientific validation of its anticancer activity. This study therefore evaluated the antiproliferative activity of E. ingens on the DU-145 human PCa cell line. The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) bio-assay was used to determine the antiproliferative activity of E. ingens ethyl acetate-, and water-fractions. Only the ethyl acetate fraction was considered to be active. It inhibited DU-145 cell growth selectively without toxicity to non-cancerous Vero E6 cells. The IC50 was 9.71 ± 0.4 µg/ml and had a selectivity index of 8.26, indicating promising efficacy and cellular safety. Qualitative phytochemical screening identified the presence of phenols, terpenoids, flavonoids, tannins, sterols, and saponins in the ethyl acetate fraction. An additional 18 chemical compounds with potential synergistic roles were identified through gas chromatography-mass spectrometry analysis. Network pharmacology was then employed to predict molecular targets and mechanisms of action for drug-like chemical compounds, implicating key targets such as ESR1, IL6, MMP9, CDK2, MAP2K1, AR, PRKCD, CDK1, CDC25B, and JAK2 with regulation of PI3K/Akt, MAPK, and p53 signaling pathways suggested to be the potential mechanism of action. Further, gene expression analysis of selected targets through reverse transcription-quantitative polymerase chain reaction revealed downregulation of AR and BCL2 levels, along with upregulation of p53 and caspase-3 in fraction-treated DU-145 cells compared to the 0.2 % DMSO-negative control. In conclusion, the findings of this study validate the traditional use of E. ingens in cancer management. The identified drug-like compounds, their targets, and associated signaling pathways could serve as a foundation for developing novel strategies for prostate cancer management. However, we recommend additional in vitro and in vivo studies to further substantiate these findings.