Whole-transcriptome Analysis of Fully Viable Energy Efficient Glycolytic-null Cancer Cells Established by Double Genetic Knockout of Lactate Dehydrogenase A/B or Glucose-6-Phosphate Isomerase.

摘要: Background/aim Nearly all mammalian tumors of diverse tissues are believed to be dependent on fermentative glycolysis, marked by elevated production lactic acid and expression glycolytic enzymes, most notably dehydrogenase (LDH). Therefore, there has been significant interest in developing chemotherapy drugs that selectively target various isoforms the LDH enzyme. However, considerable questions remain as consequences biological ablation or upstream targeting pathway. Materials methods In this study, we explore biochemical whole transcriptomic effects CRISPR-Cas9 gene knockout (KO) lactate dehydrogenases A B [LDHA/B double KO (DKO)] glucose-6-phosphate isomerase (GPI KO) human colon cancer cell line LS174T, using Affymetrix 2.1 ST arrays. Results The metabolic profiles corroborate relative wild type (WT), LDHA/B DKO produced no acid, minimal both KOs displayed higher mitochondrial respiration, use glucose with loss viability. These findings show a high energy efficiency measured ATP glycolysis-null cells. Next, analysis conducted 48,226 mRNA transcripts reflect 273 differentially expressed genes (DEGS) GPI clone set, 193 DEGS set 47 DEGs common clones. Glycolytic-null cells up-regulation typically associated nutrient deprivation / fasting possible fats for energy: thioredoxin interacting protein (TXNIP), 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), PPARγ coactivator 1α (PGC-1α), acetyl-CoA acyltransferase (ACAA2). Other changes non-ergometric losses "stemness", WNT signaling pathway, chemo/radiation resistance, retinoic synthesis, drug detoxification, androgen/estrogen activation, extracellular matrix reprogramming genes. Conclusion demonstrate that: 1) "Warburg effect" is dispensable, 2) LDHAB not only inconsequential viability but fosters greater energy, 3) likely ineffective without plausible combination second target.