多药耐药基因转染骨髓造血细胞移植小鼠的安全性研究

目的 探讨多药耐药基因(mdrl)转染骨髓造血细胞移植后,其在体内分布和安全性.方法 骨髓造血细胞转染mdrl基因后移植入BALB/C鼠,免疫组织化学和RT.PCR观察mdrl基因在骨髓、外周血及重要脏器的表达;监测外周血白细胞变化及重要脏器病检分析其安全性.结果 P-糖蛋白(P-gP)在骨髓表达率随时间降低,各时间组差异无统计学意义(x2=3.74,P＞0.05),在外周血表达第2月最高,为(9.95±1.84)%,各时间组差异无统计学意义(x2=2.31,P＞0.05),在重要脏器无表达;移植未转染组与移植转染组外周血白细胞计数至移植60 d后,两者差异有统计学意义(t=6.173,P＜0.05),病检除肝和肾有炎性渗出和水肿病变外,其余无异常.结论 mdrl基因在骨髓和外周血有持续较高表达,在重要脏器无表达;mdrl基因转染对造血恢复及重要脏器无明显影响.

The safety of bone marrow transplantation with mdrl gene transfection in mice

Objective To observe the distribution and safety of the foreign muhidrug resistance gene 1 ( mdrl ) in gene therapy.Methods After the bone marrow mononuclear cells which transferred with mdrl were transplanted into BABL/C,the expression of foreign mdrl in the bone marrow,peripheral blood and chief organs were detected by immunohistochemistry and RT-PCR.The safety was detected by counting peripheral white blood ceils and hematoxylin and eosin staining of chief organs.Results The expression of mdrl in bone marrow was reduced gradually along with the time on,but there was no difference between the groups (X2= 3.74,P＞0.05 ).The strongest expression of mdrl in peripheral blood was detected at the second month,and no difference between the groups (x2 = 2.31,P＞0.05 ).There was no expression of mdrl in chief organs.There was obvious difference between experiment group and control group in the number of peripheral white blood cells at 60th day ( t = 6.173,P＜0.05 ).Inflammatory exudation and edema were observed in liver and kidney.Conclusion The expression of mdrl in bone marrow and peripheral blood was high,but no expression was detected in chief organs.The mdrl transfection had no effect on the number of peripheral white blood cells and chief organs.