Anti-neoplastic effect of chicken anemia virus VP3 protein (apoptin) in Rous sarcoma virus-induced tumours in chicken
Anti-neoplastic effect of chicken anemia virus VP3 protein (apoptin) in Rous sarcoma virus-induced tumours in chicken
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DOI：
10.1099/vir.0.82085-0
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发表时间：
2006-10-01
影响因子：
3.8
通讯作者：
Sylvester, A.
中科院分区：
文献类型：
--
作者：
Natesan, Senthilkumar;Kataria, J. M.;Sylvester, A.
研究了鸡贫血病毒VP3蛋白(Apoptin)在体外对Rous肉瘤病毒(RSV)转化的鸡胚胎成纤维细胞(CEF)和RSV诱发的无特定病原体(SPF)雏鸡体内肿瘤的抑制作用。将凋亡素基因克隆到pVAX表达载体中，证实了重组载体pVAX-CAV-VP3的体外表达。使用两组SPF雏鸡，每组包含10只雏鸡。试验I、11组1日龄雏鸡接种RSV。第1组为空白对照组，第10天接种含凋亡素基因的重组载体pVAX-CAV-VP3。体外实验证实，凋亡素可诱导RSV转化的CEF细胞发生凋亡，并通过间接免疫荧光技术和吖啶橙/溴化乙锭染色观察细胞的凋亡特征。体内研究还表明，凋亡素通过瘤内递送方式诱导细胞凋亡和肿瘤消退。组织病理学和吖啶橙/溴化乙啶染色显示肿瘤细胞发生核固缩、染色质碎裂和凋亡小体形成等细胞凋亡改变。对照组肿瘤未见细胞凋亡性改变。本研究结果表明，凋亡素在体内和体外对RSV诱导的肿瘤具有抗肿瘤作用。其抗肿瘤作用是由凋亡素诱导的细胞凋亡所致。进一步改进凋亡素表达重组载体的剂量、递送方式和递送频率，有助于开发凋亡素作为抗肿瘤药物。
The anti-neoplastic effect of chicken anemia virus VP3 protein (apoptin) was investigated in vitro in Rous sarcoma virus (RSV)-transformed chicken embryo fibroblast (CEF) cells and in RSV-induced tumours of specific-pathogen-free (SPF) chicks in vivo. The apoptin gene was cloned in the pVAX expression vector and in vitro expression of the recombinant vector pVAX-CAV-VP3 was confirmed. Two groups of SPF chicks, each containing ten chicks, were used. Chicks in groups I and 11 were inoculated with RSV at 1 day old. Group I served as the control, receiving pVAX vector without insert, and group 11 received recombinant vector pVAX-CAV-VP3 containing the apoptin gene, on day 10. An in vitro study confirmed that apoptin induced apoptosis in RSV-transformed CEF cells, which was demonstrated by observation of the characteristic changes of apoptosis using the indirect immunofluorescence technique and acridine orange/ethidium bromide staining. In vivo study also indicated that apoptin induced apoptosis and caused tumour regression by an intratumoral-delivery method. Apoptotic changes, such as nuclear condensation, fragmentation of the chromatin and formation of apoptic bodies in the tumour cells, were demonstrated by histopathology and acridine orange/ethidium bromide staining. No apoptotic changes were seen in the tumours of the control group. The results of the present study showed that apoptin had an anti-neoplastic effect in vivo and in vitro in RSV-induced tumours. The anti-neoplastic effect is due to apoptin-induced apoptosis. Further improvements in the dose, delivery method and delivery frequency of the apoptin-expressing recombinant vector could help to develop apoptin as an anti-neoplastic drug.