CAJ | 학술논문

目的探讨Apaf-1基因启动子甲基化与抑凋亡蛋白Apollon在成人急性白血病(AL)发生发展中的作用.方法采用甲基化特异性PCR(MS-PCR)检测53例AL患者骨髓细胞基因启动子区域甲基化情况,RT-PCR方法检测Apaf-1 mRNA表达水平.免疫细胞化学方法检测Apollon蛋白表达情况,正常骨髓对照为10名正常人和非恶性血液病患者.结果 18例(33.9%)AL患者Apaf-1基因启动子存在异常甲基化,甲基化检测阳性患者均未检测到Apaf-1 mRNA的表达,对照组仅1份Apaf-1 mRNA表达缺失,MS-PCR检测未发现Apaf-1基因启动子的异常甲基化,AL患者Apaf-1基因甲基化阳性率高于对照组,差异有统计学意义(P＜0.05),AL患者骨髓细胞Apollon蛋白表达水平高于对照组(P＜0.05),AL患者外周血WBC＞10×109/L的患者Apaf-1基因启动子甲基化阳性率及Apollon蛋白表达水平高于WBC≤10×109/L患者,差异有统计学意义(P＜0.05),AL患者中Apaf-1基因启动子甲基化阳性率与Apollon蛋白表达呈正相关.结论 Apaf-1基因启动子的异常甲基化与抑凋亡蛋白Apollon的高表达在白血病的发生发展中可能起协同作用,共同促进了白血病的发生、发展.
목적 탐토Apaf-1기인계동자갑기화여억조망단백Apollon재성인급성백혈병(AL)발생발전중적작용.방법 채용갑기화특이성PCR(MS-PCR)검측53례AL환자골수세포기인계동자구역갑기화정황,RT-PCR방법 검측Apaf-1 mRNA표체수평.면역세포화학방법 검측Apollon단백표체정황,정상골수대조위10명정상인화비악성혈액병환자.결과 18례(33.9%)AL환자Apaf-1기인계동자존재이상갑기화,갑기화검측양성환자균미검측도Apaf-1 mRNA적표체,대조조부1빈Apaf-1 mRNA표체결실,MS-PCR검측미발현Apaf-1기인계동자적이상갑기화,AL환자Apaf-1기인갑기화양성솔고우대조조,차이유통계학의의(P＜0.05),AL환자골수세포Apollon단백표체수평고우대조조(P＜0.05),AL환자외주혈WBC＞10×109/L적환자Apaf-1기인계동자갑기화양성솔급Apollon단백표체수평고우WBC≤10×109/L환자,차이유통계학의의(P＜0.05),AL환자중Apaf-1기인계동자갑기화양성솔여Apollon단백표체정정상관.결론 Apaf-1기인계동자적이상갑기화여억조망단백Apollon적고표체재백혈병적발생발전중가능기협동작용,공동촉진료백혈병적발생、발전.
Objective To investigate the role of Apaf-1 gene promoter methylation and apoptosis inhibitor protein Apollon in pathogenesis of acute leukemia (AL) and their clinical significance. Methods Methylation specific PCR (MSP) was used to detect the methylation status of Apaf-1 gene promoter in 53 ALpatients (28 AML, 10 ALL and 15 relapsed) and 10 healthy or nonmalignant blood diseases patients as control. RT-PCR was used to detect the expression levels of Apaf-1 mRNA and immunocytochemistry to detect the expression levels of Apollon protein. Results The abnromal methylation of Apaf-1 gene promotor in ALwas 18/53 ( 33.9% ). No Apaf-1 mRNA was detected in methylation positive patients. Only one case in healthy and nonmalignant individuals was deletion of Apaf-1 mRNA expression without abnormal methylation.The positive methylation rate in AL bone marrow mononuclear cells was significantly higher than that in controls ( P ＜ 0.05 ). The expressin levels of Apollon protein in AL patients was higher than that in control ( P ＜0.05). The positive methylation ratio and Apollon protein level were higher in white blood cell count ＞ 10 ×109/L than in ≤ 10 × 109/L ( P ＜ 0.05 ). There is a positive correlaiton between positive methylation ratio and Apollon protein expression in AL patients. Conclusion Abnormal methylation of Apaf-1 gene promotor and high expression of Apollon might involved in leukemogenesis.