Expression of the gene for transforming growth factor type alpha, transferred into A431 cells by a recombinant retroviral vector

Cells of human epidermoid carcinoma A431 were used for obtaining of the cell line constantly expressing TGF-alpha. Recombinant virions were obtained by introducing the proviral DNA into PA317 cells by means of electrotransfection. A protein with the EGF-competing activity was found in a conditioned...

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Published inMolekuliarnaia biologiia Vol. 27; no. 4; p. 833
Main Authors Pogrebnoĭ, P V, Serov, S M, Klenchin, V A, Luk'ianov, A V, Garmanchuk, L V, Tarnavskiĭ, D V, Markeeva, N V, Bykorez, A I
Format Journal Article
LanguageRussian
Published Russia (Federation) 01.07.1993
Subjects
Autoradiography
Blotting, Southern
Electrophoresis, Polyacrylamide Gel
Gene Expression
Genetic Vectors
Humans
Iodine Radioisotopes
Isoelectric Focusing
Retroviridae - genetics
Transforming Growth Factor alpha - analysis
Transforming Growth Factor alpha - genetics
Tumor Cells, Cultured
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Summary:Cells of human epidermoid carcinoma A431 were used for obtaining of the cell line constantly expressing TGF-alpha. Recombinant virions were obtained by introducing the proviral DNA into PA317 cells by means of electrotransfection. A protein with the EGF-competing activity was found in a conditioned media of the chosen clone A431/1522-4. The concentration of this protein was several times higher than in a conditioned media of wild type A431 cells. By means of electrophoresis, isoelectric focusing, and immunochemical analysis it was shown that the protein is TGF-alpha. Similarly to EGF, the extracted TGF-alpha entirely displaced 125I-EGF specifically bound to receptor. TGF-alpha produced by the A431/1522 cells also stimulated autophosphorylation of the EGF receptor.
ISSN:0026-8984