Cargando…

Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.

In this paper the biochemical properties of the antigens detected by six murine monoclonal antibodies (MAbs) are described. These MAbs react selectively with the multidrug-resistant small cell lung cancer (SCLC) cell line, H69AR, compared to its sensitive parent cell line, H69 (Mirski & Cole, 19...

Descripción completa

Detalles Bibliográficos
Autores principales: Mirski, S. E., Cole, S. P.
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 1991
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1977290/
https://www.ncbi.nlm.nih.gov/pubmed/1677258
_version_ 1782135229950984192
author Mirski, S. E.
Cole, S. P.
author_facet Mirski, S. E.
Cole, S. P.
author_sort Mirski, S. E.
collection PubMed
description In this paper the biochemical properties of the antigens detected by six murine monoclonal antibodies (MAbs) are described. These MAbs react selectively with the multidrug-resistant small cell lung cancer (SCLC) cell line, H69AR, compared to its sensitive parent cell line, H69 (Mirski & Cole, 1989). Because H69AR cells do not overexpress P-glycoprotein, the antigens detected by these MAbs may be markers for non-P-glycoprotein-mediated mechanisms of resistance. We found that the 36 kDa protein precipitated by MAb 3.186 is phosphorylated and has a pI of approximately 6.7. The 55 kDa protein precipitated by MAb 3.50 is also phosphorylated and has a pI of approximately 5.7. Several observations suggest that MAbs 3.80, 3.177 and 3.187 recognise the same 47 kDa molecule and hence only MAb 3.187 was characterised further. This MAb precipitates an acidic protein which runs as a streak on isoelectric focusing gels. The 25 and 22.5 kDa cell surface proteins precipitated by MAb 2.54 both have a pI of approximately 7.6. Treatment of immunoprecipitates with glycosidase F indicated that none of the proteins detected by MAbs 2.54, 3.187, 3.50 and 3.186 have large N-linked carbohydrates. The peptide nature of the epitopes detected by MAbs 2.54 and 3.186 was unequivocally demonstrated by precipitation from in vitro translation products of H69AR RNA. The antigens detected by MAbs 3.50 and 3.187 were not detectable in immunoprecipitates of translation products but the epitopes are probably peptides because they were destroyed by boiling in sodium dodecyl sulphate. When the reaction of the MAbs with a panel of 15 paired drug-sensitive and -resistant cell lines was examined in a cell enzyme-linked immunosorbent assay, only a few resistance associated reactions were observed. Most of the reactions were either negative or not resistance-associated. When tested with three SCLC cell lines, MAb 3.187 reacted in a manner consistent with the relative resistance of the cell lines. Antigens that had similar electrophoretic mobility to those from H69AR cells were precipitated from extracts of five human cell lines of various tumour types. These data indicate that the cross-reactivities of the MAbs are due to antigens shared among the cell lines and not just the expression of common epitopes on different proteins. Resistance-associated proteins with the biochemical properties of the antigens described in this paper have not been reported previously and they remain potential markers for the as yet to be determined mechanisms of drug resistance in SCLC and other human malignancies. IMAGES:
format Text
id pubmed-1977290
institution National Center for Biotechnology Information
language English
publishDate 1991
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-19772902009-09-10 Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines. Mirski, S. E. Cole, S. P. Br J Cancer Research Article In this paper the biochemical properties of the antigens detected by six murine monoclonal antibodies (MAbs) are described. These MAbs react selectively with the multidrug-resistant small cell lung cancer (SCLC) cell line, H69AR, compared to its sensitive parent cell line, H69 (Mirski & Cole, 1989). Because H69AR cells do not overexpress P-glycoprotein, the antigens detected by these MAbs may be markers for non-P-glycoprotein-mediated mechanisms of resistance. We found that the 36 kDa protein precipitated by MAb 3.186 is phosphorylated and has a pI of approximately 6.7. The 55 kDa protein precipitated by MAb 3.50 is also phosphorylated and has a pI of approximately 5.7. Several observations suggest that MAbs 3.80, 3.177 and 3.187 recognise the same 47 kDa molecule and hence only MAb 3.187 was characterised further. This MAb precipitates an acidic protein which runs as a streak on isoelectric focusing gels. The 25 and 22.5 kDa cell surface proteins precipitated by MAb 2.54 both have a pI of approximately 7.6. Treatment of immunoprecipitates with glycosidase F indicated that none of the proteins detected by MAbs 2.54, 3.187, 3.50 and 3.186 have large N-linked carbohydrates. The peptide nature of the epitopes detected by MAbs 2.54 and 3.186 was unequivocally demonstrated by precipitation from in vitro translation products of H69AR RNA. The antigens detected by MAbs 3.50 and 3.187 were not detectable in immunoprecipitates of translation products but the epitopes are probably peptides because they were destroyed by boiling in sodium dodecyl sulphate. When the reaction of the MAbs with a panel of 15 paired drug-sensitive and -resistant cell lines was examined in a cell enzyme-linked immunosorbent assay, only a few resistance associated reactions were observed. Most of the reactions were either negative or not resistance-associated. When tested with three SCLC cell lines, MAb 3.187 reacted in a manner consistent with the relative resistance of the cell lines. Antigens that had similar electrophoretic mobility to those from H69AR cells were precipitated from extracts of five human cell lines of various tumour types. These data indicate that the cross-reactivities of the MAbs are due to antigens shared among the cell lines and not just the expression of common epitopes on different proteins. Resistance-associated proteins with the biochemical properties of the antigens described in this paper have not been reported previously and they remain potential markers for the as yet to be determined mechanisms of drug resistance in SCLC and other human malignancies. IMAGES: Nature Publishing Group 1991-07 /pmc/articles/PMC1977290/ /pubmed/1677258 Text en https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.
spellingShingle Research Article
Mirski, S. E.
Cole, S. P.
Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.
title Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.
title_full Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.
title_fullStr Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.
title_full_unstemmed Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.
title_short Multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.
title_sort multidrug resistance-associated antigens on drug-sensitive and -resistant human tumour cell lines.
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1977290/
https://www.ncbi.nlm.nih.gov/pubmed/1677258
work_keys_str_mv AT mirskise multidrugresistanceassociatedantigensondrugsensitiveandresistanthumantumourcelllines
AT colesp multidrugresistanceassociatedantigensondrugsensitiveandresistanthumantumourcelllines