Maintenance of hypomethylation status and preferential expression of exogenous humanMDR1/PGY1 gene in mouse L cells by YAC mediated transfer View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1997-07

AUTHORS

Hitoshi Kusaba, Masaharu Nakayama, Taishi Harada, Kiyoyuki Torigoe, Eric D. Green, Stephen W. Scherer, Kimitoshi Kohno, Michihiko Kuwano, Morimasa Wada

ABSTRACT

Selection of cells for resistance to vincristine or doxorubicin often induces overexpression of the multidrug resistance (MDR) genes, which encode the cell surface P-glycoproteins, as a result of gene amplification, transcriptional activation, or mRNA stabilization. The LMD1 and LMD4 cell lines were established after the transfer into mouse L cells of two independent yeast artificial chromosome clones containing 300 and 850 kb, respectively, of the human MDR locus. The human MDR1/PGY1 gene, but not the endogenous mouse mdr1a and mdr1b genes, was overexpressed as a result of gene amplification and transcriptional activation in various sublines of LMD1 and LMD4 cells selected for resistance to vincristine. Then we asked why human MDR1/PGY1 gene, but not mouse relevant gene, was expressed. Determination of the methylation status of cytosine residues at Msp I/Hap II cleavage sites (CCGG) in the promoter regions of human MDR1/PGY1 and mouse mdr1a revealed hypomethylation and hypermethylation of the human and mouse genes, respectively in LMD1, LMD4, and their vincristine-resistant derivatives. Various vincristine-resistant sublines were also established after exposure of LMD1 cells for 48 h to 5-aza-2'-deoxycytidine, an inhibitor of DNA methyltransferase. These sublines exhibited overexpression of mouse mdr1a and mdr1b, but not of human MDR1/PGY1, as well as hypomethylation of the mouse mdr1a promoter region. Thus, the selective expression of human or mouse MDR genes in this cell system appears to be related to the methylation status of the respective gene promoter regions. More... »

PAGES

259-274

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf02674417

DOI

http://dx.doi.org/10.1007/bf02674417

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1031491261

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/9542528


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