uPAR Knockout Results in a Deep Glycolytic and OXPHOS Reprogramming in Melanoma and Colon Carcinoma Cell Lines

Cells. 2020 Jan 28;9(2):308. doi: 10.3390/cells9020308.

Abstract

Urokinase Plasminogen Activator (uPA) Receptor (uPAR) is a well-known GPI-anchored three-domain membrane protein with pro-tumor roles largely shown in all the malignant tumors where it is over-expressed. Here we have exploited the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene knock out approach to investigate its role in the oxidative metabolism in human melanoma and colon cancer as the consequences of its irreversible loss. Knocking out PLAUR, a uPAR-encoding gene, in A375p, A375M6 and HCT116, which are two human melanoma and a colon carcinoma, respectively, we have observed an increased number of mitochondria in the two melanoma cell lines, while we evidenced an immature biogenesis of mitochondria in the colon carcinoma culture. Such biological diversity is, however, reflected in a significant enhancement of the mitochondrial spare respiratory capacity, fueled by an increased expression of GLS2, and in a decreased glycolysis paired with an increased secretion of lactate by all uPAR KO cells. We speculated that this discrepancy might be explained by an impaired ratio between LDHA and LDHB.

Keywords: CRISPR; colon cancer; gene-editing; melanoma; uPAR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • CRISPR-Associated Protein 9 / metabolism
  • Cell Line, Tumor
  • Cell Respiration / genetics
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / metabolism*
  • Colonic Neoplasms / ultrastructure
  • Deoxyribonuclease I / metabolism
  • Fluorescence
  • Gene Expression Regulation, Neoplastic
  • Gene Knockout Techniques*
  • Glycolysis* / genetics
  • Humans
  • Lactic Acid / metabolism
  • Melanoma / genetics
  • Melanoma / metabolism*
  • Melanoma / ultrastructure
  • Mitochondria / metabolism
  • Mitochondria / ultrastructure
  • Organelle Biogenesis
  • Oxidative Phosphorylation*
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • Receptors, Urokinase Plasminogen Activator / genetics
  • Receptors, Urokinase Plasminogen Activator / metabolism*
  • Stress, Physiological

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • Receptors, Urokinase Plasminogen Activator
  • Lactic Acid
  • CRISPR-Associated Protein 9
  • Deoxyribonuclease I