Study of glutamine dependency for cell growth in pediatric cancer cells

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We examined the effects of glutamine deprivation on glutamine transport activity in human neuroblastoma. We used a human neuroblastoma cell line (SK-N-SH) for the study. In vitro cell growth was measured in medium containing two different glutamine concentrations (DMEM+1O%FBS+2 mM Glutamine (Control) and DMIEM+1O%FBS (Glutamine deprivation)). Plasma membrane glutamine transport was measured at 0, 8, 16 and 24 hours after glutamine deprivation and its kinetic characteristics (Km and Vmax) were determined. Data (mean SD) were analyzed and compaired with Student's t-test.Glutamine transport occurred by the high affinity Na^+-dependent carrier (System ASC). CeLl growth rates were dependent on glutamine concentrations. Glutamine transport activity was significantly decreased at 8, 16, and 24 hours after glutamine deprivation. Kinetic studies demonstrated that glutamine deprivation decreased maximum transport velocity (Vmax) (Control, 13713 * 803 pmol/mg protein/mm ; Glutamine deprivation, 9553* 646, p< 0.01) without affecting transport affinity (Km).Glutamine transport activity was decreased in glutamine-deprived human neuroblastoma by the mechanism of decreasing the number of System ASC glutamine transporter. This may diminish glutamine availability and thereby influence cell growth in glutamine-deprived cells.