Steroid Compounds of Manihot Esculenta Crantz Var. Sao Pedro Petro (Tuber) and Their Cytotoxic Effects on Melanoma Cancer Cells (B16-F10)

Abstract

The pharmacological research of Manihot esculenta has stated the potential present as an interesting cytotoxic activity caused by several compounds contained in this species. However, the research on Melanoma Malignant cells (B16-F10 cancer cells) has never been studied. In this study, the steroid compounds isolated from the tuber of Sao Pedro Petro (1 variety of species M. esculenta Crantz) were studied in vitro and further evaluated using the western blot technique. Three steroid compounds, namely β-sitosterol (1), Campesterol (2), and Cholesterol (3), were obtained from the n-hexane extract of fresh Cassava tubers (Manihot esculenta Crantz). The chemical structures of compounds 1 - 3 were determined through various spectroscopic techniques such as HR-TOFMS, and NMR (1H, 13C, DEPT 135 °, HMQC, HMBC, 1H-1H COSY), along with a comparison to previously reported spectral data. Additionally, the apoptotic signaling pathway was investigated using RT-qPCR and western blot analysis. Among the 3 compounds, Campesterol (2) exhibited the highest IC50 value and demonstrated significant inhibition of B16-F10 cell proliferation by inducing apoptosis through caspase-3 activation. In conclusion, our study is the first to show that steroid compounds isolated from M. esculenta have activity against B16-F10 cells. This provides insight that steroids have the potential as chemopreventive agents to treat melanoma skin cancer. HIGHLIGHTS Three steroid compounds, namely β-sitosterol, campesterol, and cholesterol, were isolated from the n-hexane extract of fresh cassava tubers (Manihot esculenta Crantz) and campesterol exhibited the highest cytotoxic activity against B16-F10 melanoma cells, with an IC50 value of 160.4 µg/mL Campesterol induced apoptosis in B16-F10 melanoma cells through caspase-3 activation Treatment with campesterol led to noticeable alterations in cell morphology, including changes in shape, size, and other visible characteristics, further supporting its cytotoxic effects on melanoma cells The research's novelty lies in its exploration of previously unreported steroid compounds from cassava tubers, particularly campesterol, and their cytotoxic effects on melanoma cells, along with insights into the underlying mechanisms of action These findings contribute to the growing body of knowledge on natural compounds with potential therapeutic applications in cancer treatment GRAPHICAL ABSTRACT