Transcriptomics Analysis Provides Insights into the Cytotoxicity Effect by Inonotus rickii Mushroom (Agaricomycetes) Culture Exudate in Breast and Colon Cancer Cells.

Medicinal mushrooms have higher metabolite content and thus receive considerable attention as medicines. Inonotus rickii (Pat.) Reid is one of the mushrooms found in India. The anticancer effects of I. rickii mushrooms were investigated in this study using breast cancer and colon cancer cell lines. I. rickii mushrooms are known to secrete exudate, and its cytotoxic effect on breast cancer cells (MDA-MB-231) and colon cancer cells (HCT116) was determined. Transcriptomic analysis was determined to investigate the differentially regulated genes responsible for the growth and migration of cancer cells. The cancer cells namely HCT116 and MDA-MB-231 cells were treated with 20% and 10% exudate of I. rickii for cell viability and wound healing assay was done. RNA from the treated and non-treated cells were isolated and analyzed for RNA sequencing. The RNA sequence data was analyzed for functional pathway enrichment analysis, data visualization and gene set enrichment analysis. Quantitative analysis of gene expression of some gene responsible for cancer inhibition by using real-time polymerase chain reaction. The culture exudate of I. rickii showed 25% decrease in cell viability of breast cancer cell line (MDA-MB-231) and colon cancer cell line (HCT116) at 10% (v/v) and 20% (v/v) of exudate respectively compared to HEK normal cell line. At low concentrations, comparative transcriptomics was performed after treatment of exudate to MDA-MB-231 and HCT116 cancer cells. Few differentially regulated genes included genes responsible for the growth and migration of cancer cells. The exudate treatment also showed inhibition in cell migration and also in genes involved in both the cancers. Thus, I. rickii exudate possess anti-cancer property based on cell viability, cell migration and gene expression analysis. Out of ten reported metabolites by LCMS analysis from exudate, four anticancerous metabolites viz., Kysimplexini, Pterulamide, Etzionin and Communesin H. Mushrooms are known to have many nutrients and thus can be used as a part of diet. Thus, these can be used as therapeutics for breast and colon cancer without any side effects.