半定量RT-PCR检测服用甲磺酸伊马替尼慢性粒细胞性白血病患者hOCT1的表达

To Detect the Expression of hOCT1 by Semi-quantity RT-PCR in Chronic Myelogeneous Leukemia Patients Treated with Imatinib Mesylate

目的动态检测慢性粒细胞白血病(CML)患者服用伊马替尼缓解或耐药前后的hOCT1基因表达变化,初步探讨hOCT1基因表达变化与疾病缓解或伊马替尼耐药相关性。方法采用半定量RT-PCR方法对服用伊马替尼CML患者缓解或耐药前后的2个不同时点的9对(18份)标本进行了hOCT 1基因mRNA水平检测,并对半定量RT-PCR结果结合临床表现、实验室检查和细胞遗传学结果,分析其水平变化和与疾病缓解或伊马替尼耐药相关性。结果慢性期CML患者5例,第一和二次检测hOCT1相对表达量为(0.4246±0.2067)、(0.4858±0.1943),P=0.705,差异无统计学意义。对进展期CML患者4例前后两次hOCT1相对表达量比较,有3例患者hOCT1相对表达量降低,1例患者hOCT1相对表达量升高。结论半定量RT-PCR方法方便简单,可以用于检测服用伊马替尼的CML患者hOCT1基因表达及变化,或许有可能用于慢性粒细胞白血病伊马替尼反应或耐药的检测及其机制的研究。

Objective To monitor hOCT1 gene expression in CML patients treated with imatinb mesylate and observe the correlation between hOCT1 gene variation and BCR-ABL molecular response conditions. Methods By semi-quantity RT-PCR （SQ-PCR）, hOCT1 gene mRNA levels were detected from 9 pairs of （18） samples on two different time points around the course of response or drug resistance to imatinib in patients with chronic myelogenous leukemia respectively. The results of their SQ-PCRs were compared with clinical presentations, laboratory detections and cytogenetic findings thereof, to get some information on the correlation between changes of hOCT1 gene levels and disease response or resistance to imatinib. Results The relative expression levels of hOCT1 gene were（0. 4246±0. 2067）,（0. 4858±0. 1943）, with P=0. 705 and without statistical significance in both tests 1 and 2 in 5 patients with chronic myelogenous leukemia in chronic phase, respectively. The relative expression levels of hOCT1 gene between two tests increased in 3 and decreased in 1 of 4 patients with advanced stage of chronic myelogenous leukemia respectively. Conclusion This study established a relatively simple and useful method to detect hOCT1 gene expression that prob-ably can be used in the study on imatinib resistance.