目的：探讨体外培养条件下，缺乏葡萄糖(No-Glu)或谷氨酰胺(No-Gln)对黑色素瘤细胞生长和凋亡的影响。方法：分别利用MTT、流式细胞术、化学发光、Western blotting以及免疫荧光染色等方法检测No-Glu和No-Gln对人恶性黑色素瘤A375细胞初级纤毛形成、增殖、ATP含量、细胞周期以及细胞凋亡等多方面的影响。结果：在No-Glu或No-Gln培养液中培养，A375细胞的细胞增殖率显著低于对照组(正常培养液)\[（21.0±1.9)%或(46.2±9.8)% vs 100%, P<0.01或P<005\]。No-Gln培养液使A375细胞纤毛形成阳性细胞率显著高于对照组\[（16.8±2.3)% vs (6.2±1.0)%，P<0.01\]，G1期细胞减少(14.4±6.7)%（P<0.05）、S期细胞增加(75.0±1.9)%（P<0.05）。No-Glu培养液对A375细胞纤毛形成与G1和S期细胞比例均无显著影响。此外，No-Gln使A375细胞内ATP含量下调了(64.5±4.9)%（P<0.01），而No-Glu仅使ATP含量下降了(11.1±2.1)%（P>0.05）。No-Glu使A375细胞的凋亡率显著高于对照组\[（26.1±1.5)% vs (6.1±7.1)%，P<001\]，并上调促凋亡蛋白Noxa、降低抗凋亡蛋白Mcl-1的表达水平。No-Gln对细胞凋亡率、Noxa和Mcl-1 蛋白表达无明显影响。结论：No-Glu或No-Gln均可抑制A375细胞增殖，但No-Gln导致S期细胞周期阻滞、抑制ATP合成的作用更明显，而No-Glu诱导细胞凋亡的作用更强。

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<005). 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 G1 phase (P<0.05) and increased (75.0±19)% 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 G1 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.

国家自然科学基金资助项目（No.81202536）