以LacZ基因标记的大鼠微量残留白血病细胞的分布研究

Distribution of minimal residual leukemia in Brown Norway rats express LacZ gene

目的 了解微量残留白血病 (MRL)在体内的分布状态。方法 利用携带外源性标志基因LacZ的白血病细胞株LT12nl建立了BN大鼠MRL模型。应用逆转录 聚合酶链反应 (RT PCR)及巢式PCR方法 ,结合X gal染色法、细胞形态学及病理形态学方法检测BN大鼠MRL阶段白血病细胞中的特异性标志基因LacZ。结果 化疗后第 4天肱骨标本中出现阳性条带 ;第 6天股骨出现阳性条带 ;第 9天脾脏出现阳性条带 ,肱骨条带的信号明显较前增强。同期X gal检测脾脏阴性。外周血、肝脏持续阴性。结论 PCR方法可作为实验性MRL分布模式研究的有效手段之一。

Objective To investigate the distribution of minimal residual leukemia(MRL) in vivo. Methods The MRL model in BN rat was established by using LT12nl leukemia cell line carrying LacZ gene.The marker gene(LacZ)was detected by using reverse transcriptase polymerase chain reaction(RT PCR) and nested PCR,X gal staining,and cytomorphological and pathological examination. Results PCR assays showed positive result in humerus at day 4,femur at day 6,and spleen at day 9 after CTX treatment.The peak positive signal in humerus was stronger at day 9.X gal staining showed negative in spleen at day 9.PCR showed negative results in peripheral blood and liver. Conclusion PCR can be used as an effective tool to investigate the distribution pattern in experimental MRL.