G6PD缺陷诱发人黑色素瘤A375细胞凋亡机制研究

发布时间：2018-07-11 11:06

  本文选题：黑色素瘤 + GPD缺陷　； 参考：《中华肿瘤防治杂志》2014年14期

【摘要】：目的:探讨葡萄糖-6-磷酸脱氢酶(glucose-6-phosphate dehydrogenase,G6PD)缺陷对A375细胞凋亡的影响及其可能的机制。方法:以A375-WT和A375-G6PDΔ细胞为模型,用Real-time PCR检测G6PD mRNA表达,蛋白质印迹法检测G6PD、Bcl-2、Bcl-xL、STAT5和P-STAT5蛋白的表达,紫外分光光度法测定G6PD酶活性,Heochst 33342/PI双染流式细胞仪检测细胞凋亡,免疫组化观察STAT5蛋白核迁移。结果:A375-WT和A375-G6PDΔ细胞中G6PD mRNA分别为0.545±0.13和0.207±0.03,降低了62.02%,t=-5.854,P=0.028;G6PD蛋白分别为0.975±0.16和0.227±0.10,降低了76.72%,t=-21.593,P=0.002;G6PD酶的比活性分别为(0.088±0.023)和(0.024±0.008)U/mg;降低了72.23%,t=-7.390,P=0.018;A375-G6PDΔ细胞的凋亡率为(8.62±1.67)%,比A375-WT细胞的(2.37±0.78)%升高了3.64倍,t=-12.163,P=0.007;A375-G6PDΔ细胞抗凋亡蛋白Bcl-2的表达量为0.245±0.037,比A375-WT细胞的0.578±0.073降低了57.61%,t=-16.021,P=0.004;Bcl-xL的表达量为0.138±0.019,比A375-WT细胞的0.287±0.067降低了51.92%,t=-5.377,P=0.033;A375-G6PDΔ细胞的P-STAT5/STAT5比值为0.72±0.201,比A375-WT细胞的2.28±0.367降低了68.4%(P=0.004),细胞核内STAT5表达为0.051±0.012,比A375-WT细胞核的STAT5蛋白(0.093±0.018)降低了45%(P=0.007),STAT5核迁移减少。结论:转录因子STAT5磷酸化降低、活性下降是G6PD缺陷所诱发的人黑色素瘤A375细胞凋亡的重要因素之一,为黑色素瘤发生及治疗的研究提供了新的思路。
[Abstract]:Aim: to investigate the effect of glucose-6-phosphate dehydrogenase G6PD deficiency on apoptosis of A375 cells and its possible mechanism. Methods: A375-WT and A375-G6PD 螖 cells were used as models. Real-time PCR was used to detect the expression of G6PD mRNA, Western blot was used to detect the expression of Bcl-xSTAT5 and P-STAT5, and the activity of G6PD enzyme Heochst 33342PI double staining flow cytometry was used to detect apoptosis. The nuclear migration of STAT 5 protein was observed by immunohistochemistry. 缁撴灉:A375-WT鍜孉375-G6PD螖缁嗚優涓璆6PD mRNA鍒嗗埆涓，

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