论文部分内容阅读
敲减葡糖6-磷酸脱氢酶(G6PD)表达的人黑色素瘤A375细胞(A375-G6PDΔ)呈现生长增殖抑制和凋亡率升高.为明确G6PD缺陷对裸鼠体内成瘤的影响及其可能机制,用A375-WT与A375-G6PDΔ细胞制作裸鼠荷瘤模型,观察体内瘤体生长,real-time PCR、免疫组织化学染色与紫外分光光度法分别检测瘤体组织G6PD mRNA、G6PD蛋白及酶活性,Western印迹分析凋亡相关蛋白,分光光度法测定NADPH和GSH/GSSG水平.结果显示,A375-G6PDΔ细胞注射组的裸鼠成瘤时间延长,瘤体生长明显减慢,瘤体的体积与质量显著低于A375-WT细胞注射组(P<0.01);与A375-WT细胞注射组相比,A375-G6PDΔ细胞注射组的裸鼠瘤体组织中G6PD mRNA表达、G6PD阳性细胞数与G6PD活性分别降低了87.10%、77.20%与75.77%(P<0.01),G6PD、p53和Bcl-2的表达分别降低了67.92%、65.54%和62.32%(P<0.01),Fas升高了86.38%(P<0.01),NADPH和GSH/GSSG分别降低了74.37%和86.02%(P<0.01).结果提示,G6PD缺陷可能通过减少核酸等合成的原料、改变细胞内氧化还原状态及凋亡相关蛋白表达抑制裸鼠瘤体生长与增殖,这为黑色素瘤发生和治疗研究提供了新的线索.
Apoptosis of human melanoma A375 cells (A375-G6PDΔ) knocked down by glucose 6-phosphate dehydrogenase (G6PD) showed inhibition of growth and apoptosis.For clarifying the effect of G6PD deficiency on tumorigenesis in nude mice and its The mechanism of tumorigenesis was established by using A375-WT and A375-G6PDΔ cells. The growth of tumor in vivo was observed. Real-time PCR, immunohistochemistry and UV spectrophotometry were used to detect the expression of G6PD mRNA, G6PD protein and The activity of NADPH and GSH / GSSG were measured by Western blot analysis and the levels of NADPH and GSH / GSSG were determined by spectrophotometry.The results showed that the nude mice injected with A375-G6PDΔΔT cells prolonged the tumorigenicity, the tumor growth slowed down significantly, the volume of the tumor Compared with the group injected with A375-WT cells, the expression of G6PD mRNA and the number of G6PD-positive cells in the group of A375-WT cells injected with A375-G6PDΔ cells were significantly lower than those of the group injected with A375-WT cells (P <0.01) (P <0.01). The expressions of G6PD, p53 and Bcl-2 were decreased by 67.10%, 67.52% and 62.32% (P <0.01), and NADPH and GSH / GSSG decreased by 74.37% and 86.02%, respectively (P <0.01) .The results suggested that G6PD deficiency It can provide new clues for the occurrence and treatment of melanoma by reducing the synthesis of nucleic acids and other materials, changing the intracellular redox status and the expression of apoptosis related proteins in nude mice.