实时定量PCR分析B-ALL病人TCRζ链表达特点

The Feature of TCR-zeta Chain Expression in Patients with B-ALL by Real-time Quantitative PCR

分析T细胞信号传导分子TCRζ链基因在B细胞-急性淋巴细胞白血病(B-ALL)病人外周血中的表达水平。方法:采用SYBR G reenⅠ荧光定量PCR和相对定量检测15例初发B-ALL、5例完全缓解期(CR)患者和30例正常人外周血的单个核细胞的TCRζ链表达情况,以β2微球蛋白基因(β2M)作为内参,根据相对定量公式(2-ΔΔC t)分别计算B-ALL初发患者、CR期患者与正常人TCRζ链表达差异倍数等。结果:根据相对定量公式分析显示:与正常人相比,B-ALL初发患者外周血TCRζ链表达存在3种情况,即不表达(20%)、表达下降(53%)和表达上升(27%)。TCRζ链表达水平在所有初发B-ALL病人均低于CR期病人。结论:本研究首先报道B-ALL病人中TCRζ链表达水平的变化特点,TCRζ链基因表达水平以降低为主,可能与T细胞免疫缺陷有一定关系。

Objective ： To analyze the expression level of signal transduction factor T lymphocyte-TCRξ chain gene in patients with B-acute lymphoblastic leukemia （B-ALL）. Methods：Real-Time PCR with SYBR Green Ⅰ technique was used for detecting TCRξ chain expression level in peripheral blood mononuclear cells from 15 patients with untreated de novo B-ALL, 5 patients with B-ALL completely remission （CR） and 30 normal individuals. β2 - microglobulin gene （ 132 M） was used as an endogenous reference. Relative changes in TCRξ chain expression levels was used by the 2^△△Ct method among patients with B-ALL, CR patients, and normal individuals. Results： The expression level of TCRξ chain in patients with B-ALL could be divided into absent expression （ 20% ）, down expression （53%） and over expression （27%） groups. The expression level of TCRξ chain in all de novo B-ALL patients was lower than that in CR patients. Conclusions： It is to our knowledge, the first description in the expression feature of TCRξ chain in B-ALL. The expression level of TCRξ chain was down expression in the most of B-ALL patients, and might relate to cellular immuneodeficiency.