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Molecular Genetics Section, Pediatric Branch, NIH-National Cancer Institute, Bethesda, Maryland 20892
In this report we provide evidence for the activation of distinct differentiation pathways during treatment of the neuroblastoma cell line SMS-KCNR with 1 mM dibutyryl cyclic AMP (dbcAMP) and/or 5 µM retinoic acid (RA). Our results show that the adrenal gland specific gene pG2 is induced only during dbcAMP treatment, while RA induces a neuronal phenotype and expression of all neural related genes while decreasing the expression of many chromaffin related genes. Furthermore dbcAMP does not affect the DNA content distribution of SMS-KCNR cells [G1=61.8 ± 4.1% (SD); S = 20.3 ± 6.3%; G2-M = 18 ± 5.4%] despite morphological and molecular signs of cellular differentiation. Conversely, RA arrests cell growth causing a decrease in cells in the growth fraction (S + G2 + M = 15.6 ± 6.1%) and an increase in cells in G1 (G1 = 84.3 ± 5%). Using cyclic AMP and RA in combination, we found that RA inhibited expression of adrenal gland specific gene pG2 and induced a neuronal phenotype. Since dbcAMP does not cause a significant G1 block in SMS-KCNR cells we propose that this agent may be able to induce SMS-KCNR only to an intermediate stage of chromaffin differentiation in which cells retain their proliferative potential.
1 Present address: Oncogenesis Molecular Laboratory Inst., Regina Elena, Rome, Italy.
2 To whom requests for reprints should be addressed.
Received 12/16/91. Accepted 6/ 9/92.
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