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目的探讨B细胞淋巴瘤/白血病-2基因(bcl-2)在儿童急性B淋巴细胞白血病(B-ALL)的发生、发展及预后中的意义,分析反义寡核苷酸技术(ASODN)在儿童B-ALL的临床应用前景。方法选取2006年10月至2010年10月青岛大学医学院附属医院儿科初诊B-ALL患儿36例,收集骨髓标本,获取骨髓单个核细胞(BMMCs),采用实时荧光定量PCR方法检测BMMCs中bcl-2 mRNA的表达;将bcl-2-ASODN用脂质体介导转染细胞后检测儿童B-ALL白血病细胞增殖和凋亡改变;联合bcl-2-ASODN和VDLP(长春新碱、柔红霉素、门冬酰胺酶和地塞米松)化疗药物,四甲基偶氮唑盐法(MTT法)检测bcl-2-ASODN对儿童B-ALL白血病细胞体外化疗药物敏感性的影响。结果 (1)bcl-2 mRNA在B-ALL患儿中表达升高,与对照组比较差异有统计学意义(P<0.05),化疗敏感组患儿治疗后bcl-2 mRNA表达明显减低,较化疗耐药组差异有统计学意义(P<0.05);(2)bcl-2-ASODN用脂质体介导转染后白血病细胞增殖减弱、凋亡增加,较对照组差异有统计学意义(P<0.05);(3)体外联合bcl-2-ASODN和VDLP化疗药物能够显著抑制儿童B-ALL白血病细胞增殖,增加儿童B-ALL白血病细胞对化疗药物敏感性(P<0.05)。结论 bcl-2在儿童B-ALL的发病机制中扮演重要角色;内源性bcl-2可能是儿童B-ALL细胞化疗耐药的重要原因之一,而bcl-2-ASODN能够增加儿童B-ALL细胞体外对化疗药物的敏感性。
Objective To investigate the significance of bcl-2 gene in the pathogenesis, development and prognosis of childhood acute B-cell lymphocytic leukemia (B-ALL). To investigate the significance of antisense oligonucleotide technology (ASODN) Clinical application of B-ALL in children. Methods From October 2006 to October 2010, 36 children with newly diagnosed B-ALL in pediatric department of Medical College Affiliated Hospital of Qingdao University were enrolled. Bone marrow samples were collected to obtain bone marrow mononuclear cells (BMMCs). Real-time fluorescent quantitative PCR was used to detect the expression of bcl 2 mRNA was detected by flow cytometry. The proliferation and apoptosis of B-ALL leukemic cells were detected by liposome-mediated transfection of bcl-2-ASODN. Combined with bcl-2-ASODN and VDLP (vincristine, Mycophenolate, asparaginase and dexamethasone) chemotherapy, and MTT assay were used to detect the effect of bcl-2-ASODN on chemosensitivity of B-ALL leukemia cells in vitro. Results (1) The expression of bcl-2 mRNA in B-ALL patients was significantly higher than that in the control group (P <0.05). The bcl-2 mRNA expression in the sensitive group was significantly lower than that in the control group (2) The proliferation and apoptosis of bcl-2-ASODN transfected leukemia cells were significantly decreased after lipofectamine mediated transfection compared with the control group (P <0.05) (P <0.05) P <0.05). (3) Combination chemotherapy of bcl-2-ASODN and VDLP in vitro could significantly inhibit the proliferation of B-ALL leukemia cells in children and increase the chemosensitivity of B-ALL leukemia cells in children (P <0.05). Conclusion bcl-2 plays an important role in the pathogenesis of childhood B-ALL. Endogenous bcl-2 may be one of the important reasons for chemoresistance of B-ALL cells in children, and bcl-2-ASODN can increase the expression of B- ALL cells in vitro chemosensitivity.