Knock-out of 5-lipoxygenase in overexpressing tumor cells-consequences on gene expression and cellular function

Cancer Gene Ther. 2023 Jan;30(1):108-123. doi: 10.1038/s41417-022-00531-9. Epub 2022 Sep 16.

Abstract

5-Lipoxygenase (5-LO), the central enzyme in the biosynthesis of leukotrienes, is frequently expressed in human solid malignancies even though the enzyme is not present in the corresponding healthy tissues. There is little knowledge on the consequences of this expression for the tumor cells regarding gene expression and cellular function. We established a knockout (KO) of 5-LO in different cancer cell lines (HCT-116, HT-29, U-2 OS) and studied the consequences on global gene expression using next generation sequencing. Furthermore, cell viability, proliferation, migration and multicellular tumor spheroid (MCTS) formation were studied in these cells. Our results show that 5-LO influences the gene expression and cancer cell function in a cell type-dependent manner. The enzyme affected genes involved in cell adhesion, extracellular matrix formation, G protein signaling and cytoskeleton organization. Furthermore, absence of 5-LO elevated TGFβ2 expression in HCT-116 cells while MCP-1, fractalkine and platelet-derived growth factor expression was attenuated in U-2 OS cells suggesting that tumor cell-derived 5-LO shapes the tumor microenvironment. In line with the gene expression data, KO of 5-LO had an impact on cell proliferation, motility and MCTS formation. Interestingly, pharmacological inhibition of 5-LO only partly mimicked the KO suggesting that also noncanonical functions are involved.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arachidonate 15-Lipoxygenase / genetics
  • Arachidonate 15-Lipoxygenase / metabolism
  • Arachidonate 5-Lipoxygenase* / genetics
  • Arachidonate 5-Lipoxygenase* / metabolism
  • Cell Line
  • Gene Expression
  • Humans
  • Neoplasms* / genetics
  • Signal Transduction
  • Tumor Microenvironment

Substances

  • Arachidonate 5-Lipoxygenase
  • Arachidonate 15-Lipoxygenase