Anti-proliferative effects of unmodified antisense oligodeoxynucleotides targeted against c-raf mRNA: use of poly (lysine/serine) copolymers or cationic lipopolyamines.
1. It is now known that nuclease-resistant phosphorothioate antisense oligodeoxynucleotides (ODN) have some actions that are unrelated to antisense mechanisms. In the present study we assessed the anti-proliferative effects of phosphorothioate (PS) and phosphodiester (PO; unmodified) antisense ODN targeted against c-raf mRNA on pancreatic cancer cells in vitro, using poly (lysine/serine) copolymers conjugated with polyethylene glycol (PLSP) or cationic lipopolyamines (Transfectam) as carriers. 2. The anti-proliferative effect of the PO antisense ODN was significantly (P < 0.05) greater than that of the PS ODN, either complexed with PLSP (2 mumol/L ODN) or the Transfectam (0.5 mumol/L ODN). However, the effect of the PS or PO antisense ODN was not dependent on the antisense sequence. The c-raf mRNA levels, assessed by reverse transcription-polymerase chain reaction, were obviously reduced by both PO and PS antisense ODN compared with mismatched ODN when complexed with the Transfectam (1 mumol/L ODN). 3. Although the anti-proliferative effects were mainly unrelated to antisense mechanisms, unmodified antisense ODN complexed with some carriers could be used as anti-tumour agents considering that synthetic carriers can be modified to improve functions, such as delivery.
Aoki
Y
Y
Second Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Japan. yaoki55@med.shinshu-u.ac.jp
Kawa
S
S
Karasawa
Y
Y
Horiuchi
A
A
Kiyosawa
K
K
D002412:Cations; D002455:Cell Division / Q000187:drug effects; D004337:Drug Carriers; D005998:Glycine / Q000008:administration & dosage / Q000031:analogs & derivatives; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology*; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000473:pathology; D010455:Peptides / Q000008:administration & dosage; D010592:Pharmaceutic Aids / Q000494:pharmacology; D011073:Polyamines / Q000008:administration & dosage*; D011092:Polyethylene Glycols / Q000008:administration & dosage; D011107:Polylysine / Q000008:administration & dosage; D019908:Proto-Oncogene Proteins c-raf / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D013096:Spermine / Q000008:administration & dosage / Q000031:analogs & derivatives; D013873:Thionucleotides / Q000008:administration & dosage / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D002412:Cations; D004337:Drug Carriers; D016376:Oligonucleotides, Antisense; D010455:Peptides; D010592:Pharmaceutic Aids; D011073:Polyamines; D012333:RNA, Messenger; D013873:Thionucleotides; C061134:dioctadecylamidoglycylspermine; D011107:Polylysine; C027794:polyserine; D013096:Spermine; D011092:Polyethylene Glycols; D019908:Proto-Oncogene Proteins c-raf; D005998:Glycine