An antisense oligodeoxynucleotide that depletes RI alpha subunit of cyclic AMP-dependent protein kinase induces growth inhibition in human cancer cells.
Enhanced expression of the RI alpha subunit of cyclic AMP-dependent protein kinase type I has been correlated with cancer cell growth. We provide evidence that RI alpha is a growth-inducing protein that may be essential for neoplastic cell growth. Human colon, breast, and gastric carcinoma and neuroblastoma cell lines exposed to a 21-mer human RI alpha antisense phosphorothioate oligodeoxynucleotide (S-oligodeoxynucleotide) exhibited growth inhibition with no sign of cytotoxicity. Mismatched sequence (random) S-oligodeoxynucleotides of the same length exhibited no effect. The growth inhibitory effect of RI alpha antisense oligomer correlated with a decrease in the RI alpha mRNA and protein levels and with an increase in RII beta (the regulatory subunit of protein kinase type II) expression. The growth inhibition was abolished, however, when cells were exposed simultaneously to both RI alpha and RII beta antisense S-oligodeoxynucleotides. The RII beta antisense S-oligodeoxynucleotide alone, exhibiting suppression of RII beta along with enhancement of RI alpha expression, led to slight stimulation of cell growth. These results demonstrate that two isoforms of cyclic AMP receptor proteins, RI alpha and RII beta, are reciprocally related in the growth control of cancer cells and that the RI alpha antisense oligodeoxynucleotide, which efficiently depletes the growth stimulatory RI alpha, is a powerful biological tool toward suppression of malignancy.
Yokozaki
H
H
Cellular Biochemistry Section, National Cancer Institute, Bethesda, Maryland 20892.
Budillon
A
A
Tortora
G
G
Meissner
S
S
Beaucage
S L
SL
Miki
K
K
Cho-Chung
Y S
YS
D000595:Amino Acid Sequence; D001943:Breast Neoplasms / Q000201:enzymology / Q000473:pathology; D002455:Cell Division / Q000187:drug effects; D003110:Colonic Neoplasms / Q000201:enzymology / Q000473:pathology; D015536:Down-Regulation; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000201:enzymology / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000737:chemistry / Q000187:drug effects*; D012333:RNA, Messenger / Q000032:analysis*; D011953:Receptors, Cyclic AMP / Q000187:drug effects* / Q000378:metabolism; D013274:Stomach Neoplasms / Q000201:enzymology / Q000473:pathology; D014407:Tumor Cells, Cultured
D016376:Oligonucleotides, Antisense; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D011953:Receptors, Cyclic AMP; D011494:Protein Kinases