DNMT3A 低表达与儿童 B-ALL 的不良预后相关，并使白血病细胞对柔红霉素产生耐药性。

DNMT3A low-expression is correlated to poor prognosis in childhood B-ALL and confers resistance to daunorubicin on leukemic cells.

机构信息

Laboratory of Hematologic Diseases, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing, China.

Beijing Key Laboratory of Pediatric Hematology-Oncology, Beijing, China.

出版信息

BMC Cancer. 2023 Mar 18;23(1):255. doi: 10.1186/s12885-023-10724-6.

BACKGROUND

Little is known about DNMT3A expression and its prognostic significance in childhood B cell acute lymphoblastic leukemia (B-ALL).

METHODS

We determined DNMT3A mRNA expression in 102 children with B-ALL. Correlations with relapse-free survival (RFS) and common clinical characteristics were analyzed. DNMT3A was stably knocked out by CRISPR/Cas9 gene editing technology in Reh and 697 B-ALL cell lines. Cell proliferation activity after treated with daunorubicin (DNR) was determined by CCK8 assay in DNMT3A KO Reh and 697 cell lines.

RESULTS

DNMT3A expression in B-ALL patients who were in continuous complete remission (CCR) was higher than in those who got relapse (P = 0.0111). Receiver operating characteristic curve showed prognostic significance of DNMT3A expression (P = 0.003). Low expression of DNMT3A (≤ 0.197) was significantly correlated with poor RFS (P < 0.001) in children with B-ALL. Knock-out of DNMT3A in Reh and 697 cell lines significantly increased IC50 of DNR (P = 0.0201 and 0.0022 respectively), indicating elevated resistance to DNR.

CONCLUSION

Low expression of DNMT3A associates with poor prognosis in children with B-ALL. Knock-out of DNMT3A confers resistance to DNR on leukemic cells.

背景

关于儿童 B 细胞急性淋巴细胞白血病（B-ALL）中 DNMT3A 的表达及其预后意义知之甚少。

方法

我们测定了 102 例儿童 B-ALL 患者的 DNMT3A mRNA 表达水平。分析了其与无复发生存率（RFS）和常见临床特征的相关性。我们使用 CRISPR/Cas9 基因编辑技术在 Reh 和 697 B-ALL 细胞系中稳定敲除了 DNMT3A。在 DNMT3A KO Reh 和 697 细胞系中，用 CCK8 法测定用柔红霉素（DNR）处理后的细胞增殖活性。

结果

处于持续完全缓解（CCR）的 B-ALL 患者的 DNMT3A 表达高于复发的患者（P=0.0111）。受试者工作特征曲线显示 DNMT3A 表达具有预后意义（P=0.003）。DNMT3A 低表达（≤0.197）与 B-ALL 患儿的 RFS 差显著相关（P<0.001）。在 Reh 和 697 细胞系中敲除 DNMT3A 显著增加了 DNR 的 IC50（P=0.0201 和 0.0022），表明对 DNR 的耐药性增加。