title: Influence of partial and complete glutamine-and glucose deprivation of breast-and cervical tumorigenic cell lines.
abstract: BACKGROUND
  Due to their high proliferative requirements, tumorigenic cells possess altered metabolic systems whereby cells utilize higher quantities of glutamine and glucose. These altered metabolic requirements make it of interest to investigate the effects of physiological non-tumorigenic concentrations of glucose and glutamine on tumorigenic cells since deprivation of either results in a canonical amino acid response in mammalian cell.
  
  
  METHODS
  The influence of short-term exposure of tumorigenic cells to correlating decreasing glutamine- and glucose quantities were demonstrated in a highly glycolytic metastatic breast cell line and a cervical carcinoma cell line. Thereafter, cells were propagated in medium containing typical physiological concentrations of 1 mM glutamine and 6 mM glucose for 7 days. The effects on morphology were investigated by means of polarization-optical transmitted light differential interference contrast. Flow cytometry was used to demonstrate the effects of glutamine-and glucose starvation on cell cycle progression and apoptosis induction. Fluorometrics were also conducted to investigate the effects on intrinsic apoptosis induction (mitocapture), reactive oxygen species production (2,7-dichlorofluorescein diacetate) and acidic vesicle formation (acridine orange).
  
  
  RESULTS
  Morphological data suggests that glutamine-and glucose deprivation resulted in reduced cell density and rounded cells. Glutamine-and glucose starvation also resulted in an increase in the G2M phase and a sub-G1 peak. Complete starvation of glutamine and glucose resulted in the reduction of the mitochondrial membrane potential in both cell lines with MDA-MB-231 cells more prominently affected when compared to HeLa cells. Further, starved cells could not be rescued sufficiently by propagating since cells possessed an increase in reactive oxygen species, acidic compartments and vacuole formation.
  
  
  CONCLUSIONS
  Starvation from glutamine and glucose for short periods resulted in decreased cell density, rounded cells and apoptosis induction by means of reactive oxygen species generation and mitochondrial dysfunction. In addition, the metastatic cell line reacted more prominently to glutamine-and glucose starvation due to their highly glycolytic nature. Satisfactory cellular rescue was not possible as cells demonstrated oxidative stress and depolarized mitochondrial membrane potential. This study contributes to the knowledge regarding the in vitro effects and signal transduction of glucose and/or l-glutamine deprivation in tumorigenic cell lines.
authors.affiliation: Department of Physiology, University of Pretoria, Private Bag X323, Arcadia, 0007 South Africa.
authors.forename: Michelle Helen
authors.initials: MH
authors.lastname: Visagie
authors.affiliation: Department of Physiology, University of Pretoria, Private Bag X323, Arcadia, 0007 South Africa.
authors.forename: Thandi Vuyelwa
authors.initials: TV
authors.lastname: Mqoco
authors.affiliation: Centre for Research and Continued Engineering Development, North-West University, Lynnwood Ridge, South Africa.
authors.forename: Leon
authors.initials: L
authors.lastname: Liebenberg
authors.affiliation: Centre for Research and Continued Engineering Development, North-West University, Lynnwood Ridge, South Africa.
authors.forename: Edward Henry
authors.initials: EH
authors.lastname: Mathews
authors.affiliation: Centre for Research and Continued Engineering Development, North-West University, Lynnwood Ridge, South Africa.
authors.forename: George Edward
authors.initials: GE
authors.lastname: Mathews
authors.affiliation: Department of Physiology, University of Pretoria, Private Bag X323, Arcadia, 0007 South Africa.
authors.forename: Anna Margaretha
authors.initials: AM
authors.lastname: Joubert
keywords: Apoptosis; Glucose; Glutamine; Glycolysis; Metabolism; Morphology; Reactive oxygen species