Anticancer Peptide MCP-1 Induces Ferroptosis in Liver Cancer HCCLM3 Cells by Targeting FOXM1/ALOXE3 Signal Pathway

Abstract

FOXM1 is a crucial oncogenic transcription factor involved in almost all cancer hallmark pathways across all cancer types. Our previous work had found that FOXM1 targeted peptide P201 can strongly inhibit the growth of cancer cells including the liver cancer HCCLM3 cells. In addition, by RNA-seq of HCCLM3 cells treated with MCP-1, an anticancer peptide optimized from P201, ALOXE3, a key feature of ferroptosis was significantly elevated while FOXM1 was down-regulated, we wonder if the cell death of HCCLM3 induced by MCP-1 was associated with ferroptosis. Also, the relationship between FOXM1 and ferroptosis was less understood. Hence, in this study, we explore the effect of MCP-1 on ferroptosis and establish the associations among MCP-1, FOXM1 and ALOXE3 in HCCLM3 cells. The results showed that MCP-1 can significantly induce the elevated expression of ALOXE3, while the other important ferroptosis features including GSH, GPX4,ROS and total iron in HCCLM3 cells were all expectedly regulated. Also, ferrostatin-1, a specific inhibitor for ferroptosis, can reverse the cell death of HCCLM3 cells when co-administrated with MCP-1. TCGA database hepatocellular carcinoma gene expression analysis showed that FOXM1 was negative-related to ALOXE3 and further confirmed by the results of siRNA knockdown of FOXM1 in HCCLM3 cells. Moreover, the co-expressed genes analysis for FOXM1 and ALOXE3 revealed that many of them were closely involved in the regulation of ferroptosis. Taken together, we discovered and confirmed the induction of ferroptosis by MCP-1 in liver cancer HCCLM3 cells and primarily established the associations among MCP-1, FOXM1 and ALOXE3.