Abstract |
We have recently shown, using antisense strategy, that the RII beta regulatory subunit of cAMP-dependent protein kinase is essential for cAMP-induced growth inhibition and differentiation of HL-60 human leukemia cells. We constructed a retroviral vector for RII beta (MT-RII beta) by inserting human RII beta complementary DNA into the OT1521 retroviral vector plasmid that contains an internal mouse metallothionein-1 promoter and a neomycin resistance gene. The PA317 packaging cell line was then transfected with MT-RII beta plasmid to produce the amphotrophic stock of MT-RII beta retroviral vector. The infection with MT-RII beta and treatment with CdCl2 brought about growth arrest in HL-60 human leukemia and Ki-ras-transformed NIH 3T3 clone DT cells in monolayer culture with no sign of toxicity. The growth inhibition correlated with the expression of RII beta and accompanied changes in cell morphology; cells became flat, exhibiting enlarged cytoplasm. The growth of these cells in semisolid medium (anchorage-independent growth) was almost completely suppressed. In contrast, overexpression of the RI alpha subunit of protein kinase enhanced the cell proliferation in DT cells. The MT-RII beta-infected cells exhibited an increased sensitivity toward treatment with cAMP analogues, such as 8-Cl-cAMP and N6-benzyl-cAMP, as compared with the parental noninfected cells. In MT-RII beta HL-60 cells, N6-benzyl-cAMP treatment greatly enhanced the expression of monocytic surface markers. These results suggest that the RII beta cAMP receptor, by binding to its ligand, cAMP, acts as a tumor suppressor protein exerting growth inhibition, differentiation, and reverse transformation.
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Authors | G Tortora, A Budillon, H Yokozaki, T Clair, S Pepe, G Merlo, C Rohlff, Y S Cho-Chung |
Journal | Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research
(Cell Growth Differ)
Vol. 5
Issue 7
Pg. 753-9
(Jul 1994)
ISSN: 1044-9523 [Print] United States |
PMID | 7947390
(Publication Type: Comparative Study, Journal Article)
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Chemical References |
- Cyclic AMP-Dependent Protein Kinase RIIbeta Subunit
- Isoenzymes
- PRKAR2B protein, human
- Prkar2b protein, mouse
- Recombinant Fusion Proteins
- Cadmium
- 8-Bromo Cyclic Adenosine Monophosphate
- N(6)-benzyl-cyclic adenosine 5'-monophosphate
- Metallothionein
- 8-chloro-cyclic adenosine monophosphate
- Cyclic AMP
- Cyclic AMP-Dependent Protein Kinases
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Topics |
- 3T3 Cells
- 8-Bromo Cyclic Adenosine Monophosphate
(analogs & derivatives, pharmacology)
- Allosteric Site
- Animals
- Cadmium
(pharmacology)
- Cell Differentiation
(physiology)
- Cell Division
- Cyclic AMP
(analogs & derivatives, pharmacology, physiology)
- Cyclic AMP-Dependent Protein Kinase RIIbeta Subunit
- Cyclic AMP-Dependent Protein Kinases
(chemistry, genetics, physiology)
- Gene Expression Regulation, Neoplastic
- Genetic Vectors
- Humans
- Isoenzymes
(chemistry, genetics, physiology)
- Leukemia, Promyelocytic, Acute
(pathology)
- Metallothionein
(genetics)
- Mice
- Phenotype
- Promoter Regions, Genetic
(drug effects)
- Recombinant Fusion Proteins
(metabolism)
- Retroviridae
(genetics)
- Signal Transduction
(physiology)
- Tumor Cells, Cultured
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