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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],G1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G1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。
AIM: To investigate the effects of No-Glu or No-Gln on the growth and apoptosis of melanoma cells in vitro. Methods: Primary cilia formation, proliferation, ATP content, cell cycle, and cell cycle were detected by MTT, flow cytometry, chemiluminescence, Western blotting and immunofluorescence staining in the human melanoma A375 cells Apoptosis and many other effects. Results: The cell proliferation rate of A375 cells was significantly lower than that of the control group (21.0 ± 1.9)% or (46.2 ± 9.8)% vs 100% in No-Glu or No-Gln culture medium <0.01 or P <0.05]. No-Gln culture medium significantly increased the percentage of ciliated cells in A375 cells ([(16.8 ± 2.3)% vs (6.2 ± 1.0)%, P <0.01] <0.05), S phase cells increased (75.0 ± 1.9)% (P <0.05). No-Glu culture medium had no significant effect on the formation of cilia and the ratio of G1 and S cells in A375 cells. In addition, No-Gln reduced the ATP content in A375 cells by 64.5 ± 4.9% (P <0.01), whereas No-Glu decreased the ATP content by only 11.1 ± 2.1% (P> 0.05). No-Glu significantly increased the apoptosis rate of A375 cells (26.1 ± 1.5% vs 6.1 ± 7.1%, P <0.01) and upregulated the pro-apoptotic protein Noxa and decreased the expression of anti-apoptotic protein Mcl- 1 expression level. No-Gln had no significant effect on the apoptosis rate, Noxa and Mcl-1 protein expression. Conclusion: No-Glu or No-Gln can inhibit the proliferation of A375 cells, but No-Gln lead to cell cycle arrest in S phase, inhibit the synthesis of ATP more obviously, but No-Glu can induce apoptosis more effectively.