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Archive > Volume 23 Issue 5 > 2016,23(5):626-632. DOI:10.3872/j.issn.1007-385X.2016.05.007 Prev Next
Basic Research
Depletion of glucose or glutamine affects growth and apoptosis of melanoma cell A375
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Abstract:
Objective: To explore the effect of deletion of glucose or glutamine on growth and apoptosis of melanoma cells in vitro. Methods: MTT, flow cytometry, chemiluminescence, Western blotting and immunofluorescence staining assays were respectively used to detect primary cilia formation, proliferation, ATP contents, cell cycles and apoptosis of melanoma A375 cell on which effected by deletions of glucose and glutamine. Results: Proliferation rate of the A375 cell in none-glucose (No-Glu) or none-glutamine (No-Gln) culture medium was significantly lower than that in control group (normal culture medium) (No-Glu: \[21.0±1.9\]% vs 100%, P<0.01; No-Gln: \[46.2±9.7\]% vs 100%, P<005). No-Gln culture significantly increased positive rate of cilia formation of the A375 cell (\[16.8±2.3\]% vs \[6.2±9.8\]%, P<0.01), decreased (14.4±6.4)% of the cells at G1 phase (P<0.05) and increased (75.0±19)% of the cells at S phase (P<0.05), compared with control group. No-Glu culture medium had no obvious effect on cilia formation as well as ratios of the cells at G1 and S phases of the A375 cell. In addition, No-Gln medium reduced ATP content in the A375 cell by (64.5±4.9)% (P<0.01), and No-Glu medium only reduced ATP content in the A375 cell by (11.1±2.1)% (P<0.05). No-Glu medium increased apoptosis rate of the A375 cell more highly than that in control group (\[26.1±7.1\]% vs \[6.1±1.5\]%, P<0.01), and increased expression of pro-apoptotic protein Noxa, decreased expression of anti-apoptotic protein Mcl-1. No-Gln medium had no effect on apoptosis, and expressions of Noxa and Mcl-1 proteins. Conclusion: All of No-Glu and No-Gln could inhibit proliferation of the A375 cell, but No-Gln could result in blocking cell cycle of S phase, inhibiting ATP synthesis more obviously, however No-Glu could much more induce apoptosis of the cell.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No: 81202536）
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