Objective To study the bcl-2 mRNA expression in bone marrow mononuclear cells of childhood B-ALL, investigate the effect of bcl-2-ASODN on proliferation, apoptosis and sensitivity to chemotherapy of childhood B-ALL cells in vitro, speculate the role of bcl-2 in the mechanism, development and prognosis of childhood B-ALL, and make a perspective on the clinical application of ASODN in the future. Methods Bone marrow mononuclear cells of childhood B-ALL were used. The expression of bcl-2 was detected by Real-time Quantity PCR. Bcl-2-ASODN was transferred into cells mediated with lipofectin. Twenty-four hours later, detect the expression of bcl-2 by Real-time Quantity PCR, proliferation of leukemia cell by MIT assay and, cell apoptosis with flowcytometry. Then add chemotherapy drugs to study the chemotherapy sensitvity of leukemia cells. Results The results showed that the bcl-2 mRNA expression in childhood B-ALL was higher than control, especially in chemotherapy resistant group, and the bcl-2 mRNA expression was down regulated by chemotherapy in sensitive group. Bcl-2-ASODN was able to down-regulate bcl-2 mRNA expression, induce apoptosis of B-ALL cells, and enhance the effect of chemotherapy to inhibit cell growth. Conclusion Bcl-2 pLays an important role in childhood B-ALL. The sensitivity of childhood B-ALL cells to chemotherapy is enhanced by hcl-2-ASODN, which suggests that inner bcl-2 in leukemia cells can bring about side effect of drug resistance.%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白血病细胞增殖和调亡改变;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)bc1-2 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 bc1-2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.
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