Integrated miRNA and mRNA omics reveal neferine induced autophagy-dependent ferroptosis in human endometrial cancer in vivo and in vitro

Abstract

The most frequent type of gynecological cancer, endometrial cancer, seriously jeopardizes the health of women. Lotus seed embryos contain large amounts of the compound neferine (Nef), which has demonstrated strong anticancer properties. Nevertheless, its impacts and defense mechanisms against Endometrial cancer (EC) are still unknown. In order to identify the putative anti-cancer pathways that Nef targets, we combined mRNA-seq and miRNA-seq analyses and assessed the effects of Nef on EC both in vivo and in vitro. The integrative analysis of miRNA and mRNA omics data identified autophagy and ferroptosis as the primary anti-cancer pathways targeted by Nef. Nef stimulated the production of autophagosomes and autolysosomes, so initiating cell autophagy, as demonstrated by transmission electron microscopy. Furthermore, Nef altered Fe2+ concentration, GSH, and MDA levels, promoting ferroptosis. Molecular data indicated that Nef activated the AMPK/mTOR and SLC7A11/GPX4 pathways in vitro and in vivo, inducing autophagy and ferroptosis. The use of the 3-Methyladenine (3-MA) and Ferrostatin-1 (Fer-1) reversed Nef-induced reactive oxygen species (ROS) generation and GPX4 expression levels, confirming that Nef suppresses Ishikawa cell growth through autophagy-dependent ferroptosis. These results indicate that Nef is a viable functional food for the treatment of EC, as it can dramatically promote autophagy-dependent ferroptosis in Ishikawa cells.