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In vivo electroporetic transfer of bcl‐2 antisense oligonucleotide inhibits the development of hepatocellular carcinoma in rats
Author(s) -
Baba Miyako,
Iishi Hiroyasu,
Tatsuta Masaharu
Publication year - 2000
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(20000115)85:2<260::aid-ijc18>3.0.co;2-r
Subject(s) - oligonucleotide , sense (electronics) , in vivo , hepatocellular carcinoma , microbiology and biotechnology , fluorescein isothiocyanate , biology , electroporation , oligomer restriction , transfection , apoptosis , fluorescein , cancer research , chemistry , biochemistry , dna , fluorescence , gene , physics , quantum mechanics
To investigate the potential use of a bcl‐2 antisense oligonucleotide for therapy against hepatocellular carcinoma, we examined the effects of the electroporetic transfer of a bcl‐2 antisense oligonucleotide on rat hepatocarcinogenesis induced by N ‐nitrosomorpholine (NNM). Sprague–Dawley rats were given water containing 175 mg/l NNM for 8 weeks and received intraperitoneal injections of a bcl‐2 antisense phosphorothioate oligonucleotide, a sense oligonucleotide or a scrambled sequence oligonucleotide encapsulated in empty liposomes, at a dose of 150 μg oligonucleotide/kg body weight, every 4 weeks. One hour after injection, in vivo electroporation was performed on the liver to achieve selective transfer of the oligonucleotides. By week 16, the rats that had received the bcl‐2 antisense oligonucleotide had significantly fewer and smaller precancerous liver lesions positive for glutathione‐S‐transferase (placental type), and a significantly lower incidence of hepatocellular carcinoma in the electroporation zone than rats that had received the sense or the scrambled oligonucleotides. Moreover, the bcl‐2 antisense oligonucleotide significantly increased the apoptotic indices in foci, neoplastic nodules and in hepatocellular carcinomas. The expression of bcl‐2 mRNA also decreased, and 3′‐fragments of bcl‐2mRNA produced by cleavage at the antisense target site were detected in rat liver. Mean cellular fluorescence in the liver increased with higher doses of fluorescein‐isothiocyanate‐labeled antisense or sense oligonucleotides. Our results show that the electroporetic transfer of bcl‐2 antisense oligonucleotide can inhibit rat hepatocarcinogenesis. Int. J. Cancer 85:260–266, 2000. ©2000 Wiley‐Liss, Inc.