Cell cycle inhibition in human BE-13 T cell leukemia cells by haptoglobin-related (HPR) antisense cDNA.
We have recently cloned and sequenced a human haptoglobin-related cDNA. Hpr expression was found in various tumor cell lines. To determine whether the haptoglobin related protein (hpr) affects the growth of an established T-cell leukemia cell line, an Hpr antisense expression vector that specifically reduces hpr production was constructed. The vector was transfected into BE-13 cells, an established T-cell leukemia cell line in which Hpr is expressed. Three stable clones were isolated in which hpr protein expression was reduced. These established cell lines proliferated more slowly than vector transfected cells in proportion to Hpr antisense mRNA expression and the reduction in hpr protein production. Following a BrdU pulse, flow cytometric analysis was performed to estimate the fraction of cells in S phase. Hpr antisense transfected cells contained less cells in S phase compared to vector transfected cells. Also in soft agar, cells expressing the antisense cDNA insert, formed on average at least 7-fold fewer colonies than cells transfected with the vector alone. The data suggest that Hpr inhibitors might be of therapeutic value for T-cell leukemia.
Shalitin
C
C
Department of Biology, Technion-Israel Institute of Technology, Haifa, Israel. channa@tx.technion.ac.il
Valansi
C
C
Lev
A
A
Hurwitz
C
C
Haas
M
M
D001483:Base Sequence; D002453:Cell Cycle / Q000187:drug effects*; D017931:DNA Primers; D016373:DNA, Antisense / Q000494:pharmacology*; D018076:DNA, Complementary; D005091:Exons; D015972:Gene Expression Regulation, Neoplastic; D006242:Haptoglobins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D015458:Leukemia, T-Cell; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D016373:DNA, Antisense; D018076:DNA, Complementary; D006242:Haptoglobins; D011994:Recombinant Proteins