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Expression of retroviral vectors containing the human multidrug resistance
1 cDNA in hematopoietic cells of transplanted mice
BP Sorrentino, KT McDonagh, D Woods and D Orlic
Department of Biochemistry and Experimental Hematology, St Jude Children's
Research Hospital, Memphis, TN 38105, USA.
Transfer of the human multidrug resistance 1 (MDR1) gene to hematopoietic
stem cells offers an approach to overcome the myelosuppression caused by a
number of antineoplastic drugs. This study was designed to determine the
effect of MDR1 gene transfer on overall P- glycoprotein (P-gp) expression
in murine hematopoietic cells. Mice were transplanted with bone marrow
cells infected with either of two different MDR1 retroviral vectors. A
reverse-transcriptase polymerase chain reaction-based assay was used to
quantify expression levels of both endogenous and vector-derived P-gp
encoding transcripts in hematopoietic cells of transplanted mice.
Expression of both a truncated and full-length MDR1 mRNA species was noted
in bone marrow and spleen colony cells. The truncated message resulted from
cryptic mRNA splice sites within the MDR1 cDNA and was detected with both
vectors. Full-length message levels exceeded those from the endogenous
genes in all but one case and roughly approximated that seen in the
modestly drug-resistant cell line SW620. We conclude that transfer of MDR1
retroviral vectors resulted in a significant increase in P-gp expression in
most cases; however, aberrant splicing of MDR1 transcripts can result in
reduced expression of vector-derived P-gp.
Volume 86,
Issue 2,
pp. 491-501,
07/15/1995
Copyright © 1995 by The American Society of Hematology
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