肝癌靶向SEA/CD80共表达腺病毒在小鼠体内的分布和表达

Preliminary study on distribution of hepatoma-targeting recombinant co-expression adenovirus vector of SEA and CD80 in mice

目的:研究肝癌靶向SEA/CD80共表达腺病毒(A-S-C)在小鼠体内的分布。方法:局部注射A-S-C后,采用实时定量PCR方法检测不同时间点BALB/c小鼠血液和粪便中腺病毒的含量。免疫组织化学染色检测腺病毒在BALB/c小鼠体内各组织中的分布情况。Western blot检测目的蛋白SEA和CD80在NOD/SCID小鼠种植瘤中的表达。结果:用药后BALB/c小鼠血液和粪便中腺病毒含量迅速上升,分别在第3天和第5天达到峰值,然后又迅速降低,到第14天时基本消失。免疫组化检测5型腺病毒抗原,结果发现腺病毒主要分布在小鼠的肝组织中,其他组织中均为阴性。免疫印迹结果显示目的基因SEA的表达量也在用药后第3天达到峰值,逐渐降低,至第14天降到最低水平。结论:肝癌靶向免疫基因治疗剂A-S-C在小鼠体内代谢稳定,目的基因能够在移植瘤中特异性表达。同时,初步结果显示A-S-C对正常器官无明显毒副作用。

Objective： To investigate the preclinical distribution of hepatoma - targeting recombinant co - expres-sion adenovirus vector of SEA and CD80 in mice. Methods： The mice were local injected with A - S - C. The serum and excrement concentration of adenoviruses in BALB/c mice was tested by RT - PCR in different time point. The tissue distribution of adenoviruses in BALB/c mice was detected by immunohistochemistry. The expression of SEA and CD80 was detected in tumor tissue of NOD/SCID mice by Western blot. Results ： After BALB/c mice were injected with A - S - C, the serum and excrement concentration of adenoviruses in BALB/c mice was rising rapidly, the peak appeared in the third and fifth day apart respectively, and then quickly reduced, disappeared in the 14 day. Immunohistochemical results showed that adenoviruses were mainly distributed in the liver tissue in mice,other tissue were all the negative results. Western blot results showed that the expression of SEA was up to the peak in the third day,and gradually reduce, heaven to a minimum level to 14 day. Conclusion ： The pharmacokinetic methods of A - S - C in mice was established. The results showed that A - S - C metabolic stability in mice,the tumor tissue could efficiently express SEA and CD80. Meanwhile, preliminary results show there is no obvious side effect to normal organs.