title: Adenovirus-mediated anti-K-ras ribozyme induces apoptosis and growth suppression of human pancreatic carcinoma.
abstract: Human pancreatic cancer is a lethal malignancy, and the lesions show a very high incidence of point mutations of the K-ras oncogene. These alterations can be used as potential targets for specific ribozyme (Rz)-mediated growth suppression of the cancer cells. We designed an anti-K-ras Rz against mutant K-ras gene transcripts (codon 12, GGT to GTT) and generated a recombinant adenovirus (rAd) to express the Rz (rAd/anti-K-ras Rz). More than 95% of Capan-1 human pancreatic cells were infected with rAd/anti-K-ras Rz when treated with the virus at 200 plaque-forming units/cell. The virus, rAd/anti-K-ras Rz, significantly suppressed mutant K-ras gene expression and inhibited the growth of Capan-1 cells. At 3 days postinfection, we observed maximum growth suppression of the cells, characteristic morphological changes of apoptosis such as nuclear condensation and oligonucleosomal DNA fragmentation, and suppression of bcl-2 oncoprotein. These changes were not found in control virus-infected cells. Our results indicated that the virus rAd/anti-K-ras Rz specifically down-regulated the K-ras/bcl-2 pathway and induced apoptotic changes in Capan-1 pancreatic carcinoma cells. High-efficiency adenovirus-mediated delivery of anti-K-ras Rz could become a significant gene therapy strategy against human pancreatic cancer.
authors.affiliation: Department of Pathology, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Tsuchida
authors.forename: H
authors.initials: H
authors.lastname: Kijima
authors.forename: S
authors.initials: S
authors.lastname: Hori
authors.forename: Y
authors.initials: Y
authors.lastname: Oshika
authors.forename: T
authors.initials: T
authors.lastname: Tokunaga
authors.forename: K
authors.initials: K
authors.lastname: Kawai
authors.forename: H
authors.initials: H
authors.lastname: Yamazaki
authors.forename: Y
authors.initials: Y
authors.lastname: Ueyama
authors.forename: K J
authors.initials: KJ
authors.lastname: Scanlon
authors.forename: N
authors.initials: N
authors.lastname: Tamaoki
authors.forename: M
authors.initials: M
authors.lastname: Nakamura
mesh_terms: D000256:Adenoviridae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D017209:Apoptosis / Q000235:genetics; D002474:Cell-Free System; D003001:Cloning, Molecular; D018014:Gene Transfer Techniques; D006131:Growth Inhibitors / Q000138:chemical synthesis / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D006868:Hydrolysis; D016297:Mutagenesis, Site-Directed; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy / Q000648:ultrastructure; D010957:Plasmids / Q000138:chemical synthesis / Q000494:pharmacology; D016283:Proto-Oncogene Proteins p21(ras) / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D016337:RNA, Catalytic / Q000138:chemical synthesis / Q000494:pharmacology*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D006131:Growth Inhibitors; D016337:RNA, Catalytic; D012333:RNA, Messenger; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)