Dihydrofolate reductase gene amplification-associated shift of differentiation in methotrexate-adapted HT-29 cells

doi:10.1083/jcb.115.5.1409

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Dihydrofolate reductase gene amplification-associated shift of differentiation in methotrexate-adapted HT-29 cells

T Lesuffleur, A Barbat, C Luccioni, J Beaumatin, M Clair, A Kornowski, E Dussaulx, B Dutrillaux and A Zweibaum
Unite de Recherches sur la Differenciation Cellulaire Intestinale, INSERM U178, Villejuif, France.

Postconfluent cultures of HT-29 cells form a heterogeneous multilayer of which greater than 95% of the cells are undifferentiated. In contrast, when stably adapted to normally lethal concentrations of methotrexate (10(-6)-10(-5) M), they form a monolayer of gobletlike cells (Lesuffleur et al., 1990) which secrete large quantities of mucins and display a discrete brush border with the presence of villin, dipeptidylpeptidase-IV, and carcinoembryonic antigen. When adapted to even higher concentrations of methotrexate (10(-4) and 10(-3) M) there is a shift in the pattern of differentiation from gobletlike to dome- forming absorptive-like cells. These cells still display an apical brush border which expresses villin and dipeptidylpeptidase-IV, but no longer express significant levels of mucins and carcinoembryonic antigen. This shift of differentiation coincides with a sudden amplification of the gene coding for dihydrofolate reductase and an increased activity of the enzyme.
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