实时定量PCR检测B细胞恶性肿瘤中BCL11A基因的表达水平

Quantitative detection of BCL11A gene expression in B cell malignancies by real-time RT-PCR

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.