Growth arrest and induction of apoptosis in breast cancer cells by antisense depletion of protein kinase A-RIα subunit: p53-independent mechanism of action
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The enhanced expression of the RIα subunit of cyclic AMP-dependent protein kinase type 1 (PKA-I) has been correlated with cancer cell growth. We have investigated the effects of sequence-specific inhibition of RIα gene expression on the growth of MCF-7 human breast cancer cells. We report that RIα antisense treatment results in a reduction in RIα expression at both mRNA and protein levels and inhibition of cell growth. The growth inhibition was accompanied by changes in cell morphology, cleavage of poly(ADP-ribose) polymerase (PARP) and appearance of apoptotic nuclei. In addition, bcl-2 protein level was reduced and p53 expression increased in growth arrested cells. Interestingly, RIα antisense inhibited cell viability and induced apoptosis in the absence of p53, suggesting that these actions of RIα antisense are exerted independent of p53. In contrast, two- and four-base mismatched control oligonucleotides had no effect on either cell growth or morphology. These results demonstrate that the RIα antisense, which efficiently depletes the growth stimulatory molecule RIα, induces cell differentiation and apoptosis, providing a new approach to combat breast cancer cell growth.
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- 1.Krebs EG, Beavo JA: Phosphorylation-dephosphorylation of enzymes. Ann Rev Biochem 88: 923–939, 1979Google Scholar
- 2.Beebe SJ, Corbin JD: Cyclic nucleotide-dependent protein kinases. In: PD Boyer, EG Krebs (eds). The Enzymes: Control by Phosphorylation, Part A, Vol. 17. Academic Press, New York, 1986, pp 43–111Google Scholar
- 4.Levy FO, Øyen O, Sandberg M, Tasken K, Eskild W, Hansson V, Jahnsen T: Molecular cloning, complementary deoxyribonucleic structure and predicted full-length amino aid sequence of the hormone-inducible regulatory subunit of 3¢,5¢-cyclic adenosine monophosphate-dependent protein kinase from human testis. Mol Endocrinol 2: 1364–1373, 1988PubMedGoogle Scholar
- 15.Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG, Earnshaw WC: Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature (London) 371: 346–347, 1994Google Scholar
- 23.Shibahara S, Mukai S, Morisawa H, Nakashima H, Kobayashi S, Yamamoto N: Inhibition of human immunodeficiency virus (HIV-1) replication by synthetic oligo-RNA derivatives. Nuc Acids Res 17: 239–252, 1989Google Scholar
- 24.Metelev V, Lisziewiez J, Agrawal S: Study of antisense oligonucleotide phosphorothioates containing segments of oligodeoxynucleotides and 2′-O-methyloligoribonucleotides. Bioorg Med Chem Lett 4: 2929–2934, 1994Google Scholar
- 28.Wyllie AH: Cell death: The significance of apoptosis. Int Rev Cytol 5: 131–136, 1980Google Scholar
- 32.Vaux DL, Cory S, Adams JM: Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature 335: 440–442, 1988Google Scholar
- 34.Miyashita T, Reed JC: Bcl-2 oncoprotein blocks chemotherapy-induced apoptosis in a human leukemia cell line. Blood 11: 151–157, 1993Google Scholar
- 38.Pepe S, Sandomenico C, Damiano V, Ruggiero A, Nesterova M, Bianco AR, Tortora G, Cho-Chung YS: Role of protein kinase A type I (PKAI) in the regulation of HL-60 human promyelocytic leukemia cell cycle kinetics. Proc Am Assoc Cancer Res 37: (abstr) 33, 1996Google Scholar
- 39.Cvijic ME, Yang W-L, Chin K-V: Cisplatin resistance in cAMPdependent protein kinase mutants. Pharmacol Ther, 1997 (in press)Google Scholar