癌症靶向基因-病毒ZD55-XAF1抗肝癌移植瘤的生长及其安全性研究  

The Study on Anticancer Effect of Targeting Gene-Virus ZD55-XAF1 in Liver Cancer Xenograft of Mice and Its Safety

作　　者：马步云[1] 何婉婉[1] 周立[1] 王毅刚[1] 

机构地区：[1]浙江理工大学生命科学学院新元医学与生物技术研究所,杭州310018

出　　处：《中国生物工程杂志》2014年第1期15-20,共6页China Biotechnology

基　　金：国家自然科学基金(81272687);浙江省自然科学基金(Z13H160020)资助项目

摘　　要：目的:探讨溶瘤腺病毒(ZD55-gene)作为载体携带外源抗癌基因(XAF1)抗肝癌移植瘤的生长及其安全性。方法:抽提溶瘤腺病毒ZD55-XAF1的基因组DNA,PCR扩增鉴定病毒;细菌平板培养和支原体检测试剂盒检测细胞有无细菌、支原体污染;通过荷瘤小鼠动物实验,观察溶瘤腺病毒ZD55-XAF1对肝癌移植瘤生长的抑制、小鼠的临床反应指标、血清肝毒性指标、各脏器组织中的病毒残留分布及病理切片观察。结果:细胞培养过程无细菌和支原体污染;较对照组,受试小鼠血清肝酶AST活性上升(P<0.05),而ALT和ALP活性基本无变化(P>0.05);PCR检测各脏器均有病毒基因组DNA存在;HE染色显示受试小鼠各脏器具有不同程度的损伤,病毒处理对肿瘤细胞具有明显的杀伤效果,而受试小鼠的临床反应并无明显异常。结论:溶瘤腺病毒ZD55-XAF1能够抑制肿瘤生长,杀死肿瘤细胞,对小鼠血清肝酶活性影响较小而对各脏器有不同程度的轻微损伤,作为癌症基因治疗载体有潜在的应用价值但其安全性还有待提高。Purpose：To investigate the safety in oncolytic adenovirus （ZD55-gene） as a vector, introducing antitumor gene （ XAF1 ） at the process of cancer therapy. Methods ： Genomic DNA of oncolytic adenovirus ZD55- XAF1 was extracted and PCR was used to identify the ZD55-XAF1;To measure the bacteria and inycoplasma contamination during cell culture, supernatant was cultivated in LB solid medium and mycoplasma was tested by detection kits; After intravenous injection of ZD55-XAF1, several indicators were evaluated, including antitumor activity of ZD55-XAF1, clinical responses of the mice, levels of serum enzymes ALT, AST and ALP, residual and distribution of the virus in tissues, HE staining of various organs. Results ： No contamination was found during cell culture, which were used for virus amplification. Except the AST, the levels of serum enzymes were normal（ P 〉 0.05）. Residual and distribution of the virus in tissue stayed normal. But HE staining showed damage on mice organs and tumor was suppressed. Conclusion：Oncolytic adenovirus ZD55-XAF1 can effectively suppress tumor growth and viability of tumor cells with little impact on serum enzymes . But hematoxylin and eosin （HE） staining showed different degrees of damage on mice organs. Oncolytic adenovirus as a vector for cancer gene therapy has potential applied value but its safety needs to be improved.

关 键 词：溶瘤腺病毒 基因治疗载体 安全性 

分 类 号：Q813[生物学—生物工程]