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目的:研究成人急性白血病患者p16基因纯合缺失和甲基化变化与其表达之间的关系,探讨其在成人急性白血病发生发展中的生物学意义。方法:分别用多重聚合酶联反应(PCR)技术及甲基化敏感限制性内切酶-PCR技术检测了71例成人急性白血病患者DNA中p16基因纯合缺失及甲基化情况;应用原位杂交及免疫化学技术检测p16mRNA及p16蛋白表达。结果:71例成人急性白血病患者中,2例检测出p16基因纯合缺失。在无p16基因纯合缺失的病例中,5例急性淋巴细胞白血病(5/26)和9例急性髓性白血病(9/43)患者测出p16基因甲基化。p16 mRNA及P16蛋白的阳性率分别为70.4％(50/71例)、61.9％(44/71例)。结论:在成人急性白血病的发生发展中,p16基因甲基化比p16基因纯合缺失更具有意义;p16基因表达异常与成人急性白血病的发生发展密切相关。
Objective: To study the relationship between the changes of p16 gene homozygosity deletion and methylation in adult patients with acute leukemia and its expression, and to explore its biological significance in the development of adult acute leukemia. Methods: The homozygous deletion and methylation of p16 gene in DNA of 71 adult patients with acute leukemia were detected by multiplex polymerase chain reaction (PCR) and methylation-sensitive restriction endonuclease-PCR (PCR-RFLP) respectively. Hybridization and immunochemical techniques were used to detect p16 mRNA and p16 protein expression. RESULTS: Of the 71 adult patients with acute leukemia, 2 had a loss of homozygous p16 gene. In cases without homozygous deletion of the p16 gene, p16 gene methylation was detected in 5 patients with acute lymphoblastic leukemia (5/26) and 9 with acute myeloid leukemia (9/43). The positive rates of p16 mRNA and P16 protein were 70.4% (50/71 cases) and 61.9% (44/71 cases) respectively. Conclusion: The methylation of p16 gene is more meaningful than the homozygous deletion of p16 gene in the development of adult acute leukemia. The abnormal expression of p16 gene is closely related to the occurrence and development of adult acute leukemia.