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

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(P0.05),而ALT和ALP活性基本无变化(P0.05);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。

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

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 mycoplasma 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.