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目的 探讨儿童急性淋巴细胞白血病 (ALL)发生及发展的机制。方法 应用RT PCR技术检测 61个ALL细胞系和 53例儿童ALL患者的p73基因mRNA表达情况 ,并结合患者的临床资料进行分析。应用DNA甲基化酶谱测定、亚硫酸氢钠修饰的PCR和PCR产物测序等技术对 61个ALL细胞系p73基因第 1外显子的甲基化进行检测。结果 61个ALL细胞系中p73mRNA阴性表达率为31 1 % (61个中 1 9个 ) ;53例原发性儿童ALL患者中 ,p73mRNA阴性表达率为 2 6 .4 % (53例中 1 4例 ) ,p73mRNA阴性表达与ALL无病生存期、总生存期的降低有明显关系。 61个ALL细胞系中 ,39.3 %存在p73基因的高甲基化。正常淋巴细胞和不存在p73基因高甲基化的细胞系表达p73mRNA ,大多数高甲基化的细胞系不表达p73mRNA。结论 儿童ALL患者中p73mRNA存在较高的阴性表达 ,其主要机制为p73基因高甲基化 ,p73基因失活在ALL的发生及发展中起重要作用 ,p73mRNA检测对判断儿童ALL患者的预后有一定临床意义
Objective To investigate the mechanism of the occurrence and development of childhood acute lymphoblastic leukemia (ALL). Methods RT-PCR was used to detect the mRNA expression of p73 in 61 ALL cell lines and 53 pediatric ALL patients, and the clinical data were analyzed. Methylation of exon 1 of p73 gene in 61 ALL cell lines was detected by DNA methylation zymography, sodium bisulfite modified PCR and PCR product sequencing. Results The negative rates of p73 mRNA expression in 61 ALL cell lines were 31.1% (19 out of 61). The negative rate of p73 mRNA expression in 53 children with primary childhood ALL was 26.4% (14 out of 53 Cases), p73mRNA negative expression and ALL disease-free survival, the overall survival decreased significantly. Among 61 ALL cell lines, 39.3% had hypermethylation of p73 gene. Normal lymphocytes and cell lines in the absence of hypermethylation of the p73 gene express p73 mRNA, and most hypermethylated cell lines do not express p73 mRNA. Conclusion There is a high negative expression of p73mRNA in children with ALL. The main mechanism is p73 hypermethylation, p73 gene inactivation plays an important role in the occurrence and development of ALL. The detection of p73mRNA has some clinical significance in judging the prognosis of children with ALL