Lee H, Kim J, Lee B, Chang J W, Ahn J, Park J O, Choi J, Yun C O, Kim B S, Kim J H
Yonsei Cancer Center, Yonsei University College of Medicine, Seoul, Korea.
Int J Cancer. 2000 Nov 1;88(3):454-63. doi: 10.1002/1097-0215(20001101)88:3<454::aid-ijc19>3.0.co;2-t.
YKL-1, E1B 55 kDa-deleted recombinant adenovirus vector, capable of harboring a transgene casette of up to 4.9 kb, was newly constructed by reintroducing E1A and E1B 19 kDa into E1/E3-deleted adenoviral vector with a homologous recombination in E. coli. Virus replication and cytotoxicity were dramatically attenuated in all 3 different types of normal human cells. In contrast, YKL-1 efficiently replicated and induced cytotoxicity in most cancer cells, especially Hep3B and C33A cells with an inactivating p53 mutation. However, both H460 and HepG2 exhibited intermediate sensitivity to YKL-1, which was between that of Hep3B or C33A and normal human cells. The YKL-1 and DNA damaging agent, camptothecin effectively induced p53 in H460 and HepG2 as well as in normal cells. Furthermore, YKL-1 effectively prohibited both Hep3B and C33A tumor growth in nu/nu mice in a dose-dependent manner. H/E staining and TUNEL assay indicated a largely distributed necrotic area and apoptosis on its periphery. This study, therefore, indicates that YKL-1, possesses promising potential as an oncolytic adenoviral vector, which acts partially in a p53-dependent manner.
YKL-1是一种E1B 55 kDa缺失的重组腺病毒载体，能够容纳长达4.9 kb的转基因盒，它是通过在大肠杆菌中进行同源重组，将E1A和E1B 19 kDa重新导入E1/E3缺失的腺病毒载体中而新构建的。在所有3种不同类型的正常人细胞中，病毒复制和细胞毒性均显著减弱。相比之下，YKL-1在大多数癌细胞中能有效复制并诱导细胞毒性，尤其是在具有失活p53突变的Hep3B和C33A细胞中。然而，H460和HepG2对YKL-1表现出中等敏感性，介于Hep3B或C33A与正常人细胞之间。YKL-1与DNA损伤剂喜树碱能有效诱导H460和HepG2以及正常细胞中的p53。此外，YKL-1能以剂量依赖的方式有效抑制nu/nu小鼠体内Hep3B和C33A肿瘤的生长。苏木精-伊红染色和TUNEL检测表明，坏死区域分布广泛，其周边有凋亡现象。因此，本研究表明，YKL-1作为一种溶瘤腺病毒载体具有广阔的应用前景，其作用部分依赖于p53。
