本文選題：黑色素瘤 + GPD缺陷��； 參考：《中華腫瘤防治雜志》2014年14期

【摘要】：目的:探討葡萄糖-6-磷酸脫氫酶(glucose-6-phosphate dehydrogenase,G6PD)缺陷對A375細胞凋亡的影響及其可能的機制。方法:以A375-WT和A375-G6PDΔ細胞為模型,用Real-time PCR檢測G6PD mRNA表達,蛋白質(zhì)印跡法檢測G6PD、Bcl-2、Bcl-xL、STAT5和P-STAT5蛋白的表達,紫外分光光度法測定G6PD酶活性,Heochst 33342/PI雙染流式細胞儀檢測細胞凋亡,免疫組化觀察STAT5蛋白核遷移。結果:A375-WT和A375-G6PDΔ細胞中G6PD mRNA分別為0.545±0.13和0.207±0.03,降低了62.02%,t=-5.854,P=0.028;G6PD蛋白分別為0.975±0.16和0.227±0.10,降低了76.72%,t=-21.593,P=0.002;G6PD酶的比活性分別為(0.088±0.023)和(0.024±0.008)U/mg;降低了72.23%,t=-7.390,P=0.018;A375-G6PDΔ細胞的凋亡率為(8.62±1.67)%,比A375-WT細胞的(2.37±0.78)%升高了3.64倍,t=-12.163,P=0.007;A375-G6PDΔ細胞抗凋亡蛋白Bcl-2的表達量為0.245±0.037,比A375-WT細胞的0.578±0.073降低了57.61%,t=-16.021,P=0.004;Bcl-xL的表達量為0.138±0.019,比A375-WT細胞的0.287±0.067降低了51.92%,t=-5.377,P=0.033;A375-G6PDΔ細胞的P-STAT5/STAT5比值為0.72±0.201,比A375-WT細胞的2.28±0.367降低了68.4%(P=0.004),細胞核內(nèi)STAT5表達為0.051±0.012,比A375-WT細胞核的STAT5蛋白(0.093±0.018)降低了45%(P=0.007),STAT5核遷移減少。結論:轉錄因子STAT5磷酸化降低、活性下降是G6PD缺陷所誘發(fā)的人黑色素瘤A375細胞凋亡的重要因素之一,為黑色素瘤發(fā)生及治療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. 緇撴灉:A375-WT鍜孉375-G6PD螖緇嗚優(yōu)涓璆6PD mRNA鍒嗗埆涓，

本文編號：2114962