Retinoic acid receptor expression vector inhibits differentiation of F9 embryonal carcinoma cells.
Expression vectors have been constructed for a region of the human retinoic acid receptor-alpha (hRAR-alpha) and transferred into F9 embryonal carcinoma (EC) cells. When the vectors are overexpressed in F9 cells, clones can be selected for resistance to retinoic acid-induced differentiation. This effect is obtained even when the hRAR-alpha region is expressed as a beta-galactosidase fusion protein. Using the beta-galactosidase component of the fusion protein as a marker, overexpression of the fusion protein has been correlated with the retinoic acid-resistance effect. The clones resistant to retinoic acid no longer exhibit the normal retinoic acid induction of endo B cytokeratin, laminin B-1, and tissue plasminogen activator mRNAs observed with normal F9 cells. Retinoic acid induction of type IV alpha-1 collagen and Hox-1.3 RNAs is observed with these clones. When transfected with a thyroid receptor DNA-binding sequence (TRE)/thymidine kinase promoter/luciferase construct, the retinoic acid-resistant clones do not yield the same retinoic acid-induced level of luciferase obtained with F9 cells. It is hypothesized that the RAR vectors are interfering with endogenous RAR(s) in a dominant-negative manner to inhibit retinoic acid-induced differentiation of F9 EC cells.
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- Tretinoin
- Transfection
- Recombinant Fusion Proteins
- Receptors, Retinoic Acid
- Phenotype
- Neoplastic Stem Cells
- Humans
- Genetic Vectors
- Gene Expression
- Embryonal Carcinoma Stem Cells
Citation
Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- Tretinoin
- Transfection
- Recombinant Fusion Proteins
- Receptors, Retinoic Acid
- Phenotype
- Neoplastic Stem Cells
- Humans
- Genetic Vectors
- Gene Expression
- Embryonal Carcinoma Stem Cells