Identification of candidate genes induced by retinoic acid in embryonal carcinoma cells.
Retinoic acid (RA) induced the terminal differentiation of a human embryonal carcinoma cell line (NT2/D1) into several morphologically distinct cell types, including the postmitotic CNS neurons. Although RA has been suggested to play an important role in brain development, little is known about the molecular mechanism by which RA induces neuronal differentiation. In the present study, RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) was used to identify the transcripts in NT2/D1 cells that were differentially regulated by RA. Northern blot analysis of the differentially amplified PCR fragments revealed 11 genes that were regulated by RA. Of these, seven were up-regulated and four were down-regulated along the course of RA treatment. More importantly, four of the RA-regulated genes that were identified in the present study are novel. Our findings suggested that there are a number of RA-regulated genes that have yet to be identified. RAP-PCR provides a useful tool for studying the patterns of transcript expression during the course of RA treatment and allows the cloning of novel genes involved in the process of neuronal differentiation. Furthermore, it provides a basis for the selection of genes that are involved in the RA-induced signaling pathway in the human CNS.
Cheung
W M
WM
Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay.
Chu
A H
AH
Ip
N Y
NY
D015152:Blotting, Northern; D018236:Carcinoma, Embryonal / Q000235:genetics*; D002454:Cell Differentiation; D053938:DNA Fragmentation; D018076:DNA, Complementary / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D005796:Genes; D005821:Genetic Techniques; D006801:Humans; D009474:Neurons / Q000473:pathology; D016133:Polymerase Chain Reaction; D012334:RNA, Neoplasm / Q000235:genetics; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D018076:DNA, Complementary; D012334:RNA, Neoplasm; D014212:Tretinoin