Effect of 1-beta-D-arabino-furanosyl-cytosine (ara-C) induction of K562 cells on expression of Rh and other blood group active proteins.
K562 cells undergoing differentiation induced by 1-beta-D-arabino-furanosyl-cytosine (ara-C) were examined as a model for studying the biosynthesis and regulation of Rh and other blood group active membrane proteins. Untreated and ara-C-induced K562 cells were analysed for the expression of these proteins using monoclonal antibodies in combination with flow cytometry. The major membrane proteins glycophorins A and C remained unaltered upon induction by ara-C. The display of LFA-3 (CD58) and DAF (CD55) by uninduced K562 was one order of magnitude lower than that of the glycophorins; following ara-C treatment there was a 50% rise in LFA-3 but a modest decrease in the level of DAF expression. The expression by untreated K562 cells of Rh, Lutheran and Kell proteins as well as the Rh D antigen was low, whereas that of CD44 and band 3 protein was negligible. Following induction by ara-C the levels of Rh and Kell proteins rose up to 7- and 3.5-fold respectively, and there was an increase in RhD-antigen expression. In contrast, ara-C induction of K562 cells failed to augment their display of Lutheran, CD44 and band 3 proteins. Analysis of Rh transcripts following the purification and RT-PCR analysis of K562 mRNA showed that uninduced K562 cells contain two distinct mRNAs corresponding to Rh Ce (1.8 kb) and Rh D (3.5 kb). The apparent concentration of each mRNA increased following induction with ara-C. K562 plasma membranes also contained Rh polypeptides as determined by immunoblot analysis using anti-Rh polypeptide rabbit polyclonal sera raised to Rh synthetic peptides. A novel hybrid Rh transcript corresponding to exons 1-4 of RHD and exons 5-10 of RHCE has been cloned and sequenced from ara-C induced K562 cells, and may have arisen by general recombination between the RHD and RHCE genes.
Wiener
E
E
Department of Haematology, Imperial College School of Medicine, St Mary's Hospital, London.
Shiels
A
A
Wickramasinghe
S N
SN
Avent
N D
ND
D015152:Blotting, Northern; D003561:Cytarabine / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D020014:K562 Cells / Q000187:drug effects / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012204:Rh-Hr Blood-Group System / Q000378:metabolism*
D006454:Hemoglobins; D007166:Immunosuppressive Agents; D008565:Membrane Proteins; D012204:Rh-Hr Blood-Group System; D003561:Cytarabine