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目的探讨β-珠蛋白生成障碍性贫血(β-地贫)患者外周血单个核细胞(PBMCs)中B细胞淋巴瘤/白血病11A(BCL11A)基因的表达特点。方法入选35例β-地贫患者为病例组;40例健康体检者为健康对照组,并进行红细胞计数。提取新鲜外周血PBMCs中总RNA,反转录为cDNA,采用荧光染料(SYBR GreenⅠ)实时荧光定量PCR(RT-PCR)和相对定量分析方法,以GAPDH基因为内参,检测2组PBMCs中BCL11A基因的表达情况。采用相对定量2-△△Ct法进行mRNA相对表达量比较,不同样本之间的相对表达量(%)=2-△Ct×100%。BCL11A基因mRNA相对表达量和红细胞计数、年龄和性别的相关分析中双变量符合正态分布的计量资料采用Pearson相关分析,其他变量采用Spearman相关分析。结果 RT-PCR检测BCL11A基因mRNA在2组中的表达,健康对照组BCL11A基因表达水平是病例组的2.46倍,2组比较差异有统计学意义。病例组中BCL11A基因mRNA的相对表达量为0.31%~11.60%;健康对照组中BCL11A基因mRNA的相对表达量为1.40%~26.70%,存在明显的个体差异。BCL11A基因mRNA的相对表达量与RBC计数(r=-0.21,P=0.930)、性别(r=-0.20,P=0.842)、年龄(r=-1.15,P=0.256)均无相关性。结论β-地贫患者BCL11A基因表达下降,BCL11A基因低表达可能在γ珠蛋白基因持续表达过程中发挥了一定作用。
Objective To investigate the expression of BCL11A gene in peripheral blood mononuclear cells (PBMCs) of patients with β-thalassemia. Methods 35 cases of β-thalassemia patients were selected as the case group. Forty healthy subjects were selected as healthy control group and red blood cell count was performed. Total RNA was extracted from PBMCs of fresh peripheral blood and reverse transcribed into cDNA. Fluorescent dye (SYBR Green Ⅰ) real-time quantitative PCR (RT-PCR) and relative quantitative analysis were used to detect the BCL11A gene The expression of the situation. Relative relative expression of 2-ΔΔCt method was used to compare the relative expression amount of mRNA (%) = 2-Δ Ct × 100%. BCL11A gene mRNA relative expression and red blood cell count, age and gender-related analysis of bivariate normal distribution of measurement data using Pearson correlation analysis, the other variables using Spearman correlation analysis. Results The expression of BCL11A mRNA was detected by RT-PCR in two groups. The expression level of BCL11A in healthy control group was 2.46 times of that in case group. The difference between the two groups was statistically significant. The relative expression level of BCL11A mRNA in case group was 0.31% ~ 11.60%. The relative expression level of BCL11A mRNA in healthy control group was 1.40% ~ 26.70%, with significant individual differences. The relative expression of BCL11A mRNA was not correlated with RBC count (r = -0.21, P = 0.930), sex (r = -0.20, P = 0.842) and age (r = -1.15, P = 0.256). Conclusion The expression of BCL11A gene is decreased in β-thalassemia patients, and the low expression of BCL11A gene may play a role in the sustained expression of γ-globin gene.