Influence of partial and complete glutamine-and glucose deprivation of breast-and cervical tumorigenic cell lines

Cell & Bioscience

Anna Margaretha Joubert²  George Edward Mathews¹  Edward Henry Mathews¹  Leon Liebenberg¹  Thandi Vuyelwa Mqoco²  Michelle Helen Visagie² 

[1] Centre for Research and Continued Engineering Development, North-West University, Lynnwood Ridge, South Africa;Department of Physiology, University of Pretoria, Private Bag X323, Arcadia 0007, South Africa

关键词: Glucose;    Glutamine;    Glycolysis;    Reactive oxygen species;    Morphology;    Apoptosis;    Metabolism;   

Others  :  1223179
DOI  :  10.1186/s13578-015-0030-1

 received in 2015-05-07, accepted in 2015-06-26,  发布年份 2015

【 摘 要 】

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 G ₂ M phase and a sub-G ₁peak. 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.

Conclusion

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.

【 授权许可】

   
2015 Visagie et al.

【 图 表 】

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