Study on the Effect of Glutamine on Proliferation and Survival of Bladder Cancer T24 Cells

Author(s): Sun Ningchuan, Liang Ye, Lijiang Sun, Niu Haitao

Objective To observe the effect of glutamine (Gln) on the proliferation and survival of bladder cancer T24 cells and explore its mechanism. Methods MTT was used to detect the proliferation of T24 cells in Gln(+) and Gln(-) groups under different time gradients. The best time was selected and the Gln(+) and Gln(-) groups were analyzed at the this time. Between the Gln(+)+Don gradients groups, T24 cells were tested for cell proliferation. Survival ratio and reactive oxygen species (ROS) content of T24 bladder cancer cells in Gln(+), Gln(-) and Gln(+)+Don groups were detected by Annexin V-FITC/PI double staining and ROS kits, respectively. The Gln(-) group was used as the control group, and the ROS scavenger N-acetyl-L-cysteine (NAC) was added to the experimental group to observe the cell proliferation level and survival ratio. Results Compared with Gln(+) group, the proliferation level of T24 cells in Gln(-) group decreased at 24, 48, and 72h, and it was most obvisously at 72h. Compared with the Gln(+) group, the cell proliferation levels of the Gln(-) group and the Gln(+)+Don group were significantly decreased, the ROS level were increased, and the survival rate were decreased significiently. Versus the Gln(-) group, the levels of ROS in the Gln(-)+NAC group decreased, and the proliferation level and survival rate increased. Conclusion Gln deficiency can inhibit the proliferation and survival of T24 cells by increasing ROS levels.

Glutamine, Bladder Cancer, Reactive Oxygen Species, Cell Proliferation, Cell Survival

1. Zhang, J., N.N. Pavlova, and C.B. Thompson, Cancer cell metabolism: the essential role of the nonessential amino acid, glutamine. Embo j, 2017. 36(10): p. 1302-1315.
2. 2Daye, D. and K.E. Wellen, Metabolic reprogramming in cancer: unraveling the role of glutamine in tumorigenesis. Semin Cell Dev Biol, 2012. 23(4): p. 362-9.
3. Fuchs, B.C. and B.P. Bode, Stressing out over survival: glutamine as an apoptotic modulator. J Surg Res, 2006. 131(1): p. 26-40.
4. Mates, J.M., et al., Glutamine and its relationship with intracellular redox status, oxidative stress and cell proliferation/death. Int J Biochem Cell Biol, 2002. 34(5): p. 439-58.
5. 5Ratnikov, B., et al., Glutamate and asparagine cataplerosis underlie glutamine addiction in melanoma. Oncotarget, 2015. 6(10): p. 7379-89.
6. Shanware, N.P., et al., Glutamine deprivation stimulates mTOR-JNK-dependent chemokine secretion. Nat Commun, 2014. 5: p. 4900.
7. van den Heuvel, A.P., et al., Analysis of glutamine dependency in non-small cell lung cancer: GLS1 splice variant GAC is essential for cancer cell growth. Cancer Biol Ther, 2012. 13(12): p. 1185-94.
8. Kim, H.M., Y.K. Lee, and J.S. Koo, Expression of glutamine metabolism-related proteins in thyroid cancer. Oncotarget, 2016. 7(33): p. 53628-53641.
9. Xiang, Y., et al., Targeted inhibition of tumor-specific glutaminase diminishes cell-autonomous tumorigenesis. J Clin Invest, 2015. 125(6): p. 2293-306.
10. Ko, Y.G., et al., Glutamine-dependent antiapoptotic interaction of human glutaminyl-tRNA synthetase with apoptosis signal-regulating kinase 1. J Biol Chem, 2001. 276(8): p. 6030-6.
11. Mates, J.M., et al., Pathways from glutamine to apoptosis. Front Biosci, 2006. 11: p. 3164-80.
12. Chen, L. and H. Cui, Targeting Glutamine Induces Apoptosis: A Cancer Therapy Approach. Int J Mol Sci, 2015. 16(9): p. 22830-55.
13. Hernandez-Davies, J.E., et al., Vemurafenib resistance reprograms melanoma cells towards glutamine dependence. J Transl Med, 2015. 13: p. 210.
14. Gross, M.I., et al., Antitumor activity of the glutaminase inhibitor CB-839 in triple-negative breast cancer. Mol Cancer Ther, 2014. 13(4): p. 890-901.
15. Wang, J.B., et al., Targeting mitochondrial glutaminase activity inhibits oncogenic transformation. Cancer Cell, 2010. 18(3): p. 207-19.
16. Gross, M.I., et al., Antitumor activity of the glutaminase inhibitor CB-839 in triple-negative breast cancer. Mol Cancer Ther, 2014. 13(4): p. 890-901.
17. Xiang, Y., et al., Targeted inhibition of tumor-specific glutaminase diminishes cell-autonomous tumorigenesis. J Clin Invest, 2015. 125(6): p. 2293-306.
18. Stalnecker, C.A., et al., Mechanism by which a recently discovered allosteric inhibitor blocks glutamine metabolism in transformed cells. Proc Natl Acad Sci U S A, 2015. 112(2): p. 394-9.