目的:建立B细胞淋巴瘤/白血病BCL11A基因的定量检测方法,并分析其在B细胞恶性肿瘤中的表达水平。方法:利用实时定量RT-PCR分析B细胞淋巴瘤(18例)、B细胞性慢性淋巴细胞白血病(B-CLL,8例)、T细胞性急性淋巴细胞白血病(T-ALL,8例)和正常对照(15例)外周血单个核细胞(PBMC)中BCL11A基因的表达水平,以甘油醛-3-磷酸脱氢酶(GAPDH)基因作为内对照。结果:B-CLL组和B细胞淋巴瘤组患者PBMC中BCL11A表达水平均明显高于正常对照(P=0.000)和T-ALL组(P=0.000);T-ALL组和正常对照组BCL11A表达水平无显著差别(P=0.084);B-CLL组和B细胞淋巴瘤组BCL11A表达水平无显著差别(P=0.776)。在B细胞淋巴瘤不同的病例中,BCL11A表达水平有较大差异,其中最小值为0.04,最大值为9.70,中位值为1.00。结论:成功建立BCL11A基因的定量检测方法。
Aim:To establish a real-time PCR quantitative detection method for B-cell lymphoma/leukemia(BCL11A) gene,and to evaluate its expression level in B cell malignancies.Methods: Real-time PCR with SYBR GreenⅠtechnology was used to detect the expression level of BCL11A gene in peripheral blood mononuclear cells from 15 normal individuals(as control) and patients with B cell lymphomia(18 cases),B-cell chronic lymphocytic leukemia(B-CLL,8 cases),T-cell acute lymphocytic leukemia(T-ALL,8 cases).Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as reference to obtain the relative expression levels of BCL11A.Results: The expression levels of BCL11A in B-CLL and B cell lymphomia was significantly higher than those in the normal control(P=0.000)and T-ALL(P=0.000).Whereas,The expression levels of BCL11A in T-ALL had no significant difference when compared with the control group(P=0.084).The expression levels of BCL11A in B-CLL had no significant difference compared with B cell lymphomia(P=0.776).In different patients of B cell lymphomia,the expression levels of BCL11A were found to be different,with the maximum value of 9.70,the minimum value 0.04.The median was found to be 1.00.Conclusion:The method for quatitation of BCL11A gene expression by Real-time PCR with SYBR GreenⅠtechnology has been established.