Fungal steroids and steroid derivatives as potential anticancer agents: A systematic review

Introduction: Cancer treatment is limited by the development of drug resistance. Therefore, new therapeutics are sought out, including fungal secondary metabolites. This study presents an overview of fungal steroids and steroid derivatives as potential anticancer agents, focusing on their extraction and cytotoxicity. Objectives: To identify fungal species producing steroidal compounds with reported cytotoxic activity, characterise main extraction trends, catalogue cytotoxically active steroids and their derivatives and evaluate their in vitro cytotoxicities across different cancer cell lines, and summarise the available evidence on their anticancer mechanisms. Methods: This systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta‑Analyses guidelines. A predefined set of keywords was used to search articles in PubMed, Taylor & Francis, and ScienceDirect. Articles were screened against predefined inclusion and exclusion criteria. Data from the included studies were then extracted and organised into tables. Results: Ten studies were included in the final analysis. The most common extraction techniques were silica gel column chromatography, high-performance liquid chromatography, and thin layer chromatography. Methanol and ethyl acetate were used as extraction solvents in seven of the ten included studies, and eight studies employed mass spectrometry, identifying a total of 20 compounds. Cytotoxicity was assessed by colorimetric assays such as the MTT assay, and the MCF7 breast cancer cell line was used in five studies. The lowest IC₅₀ values were observed for inotodiol (1.8–2.1 μg/mL after 48 h) and cholestanol glucoside (6 μM after 24 h). Inotodiol, trametenolic acid, and cholestanol glucoside also showed synergistic effects. Two studies evaluated in vivo effects in mice. Reported anticancer mechanisms included anti-inflammatory and antioxidant activity, induction of autophagy and apoptosis, and inhibition of angiogenesis. Conclusion: This review summarised the main methodological patterns that can inform future research, highlighted the lack of in vivo studies, and identified key directions for further investigation.