Shalitin C, Valansi C, Lev A, Hurwitz C, Haas M
Department of Biology, Technion-Israel Institute of Technology, Haifa, Israel.
Anticancer Res. 1998 May-Jun;18(3A):1745-50.
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.
我们最近克隆并测序了一个人触珠蛋白相关的cDNA。在多种肿瘤细胞系中发现了Hpr表达。为了确定触珠蛋白相关蛋白（hpr）是否影响已建立的T细胞白血病细胞系的生长，构建了一种能特异性降低hpr产生的Hpr反义表达载体。将该载体转染到BE - 13细胞中，BE - 13是一种已建立的、表达Hpr的T细胞白血病细胞系。分离出三个稳定克隆，其中hpr蛋白表达降低。这些已建立的细胞系与载体转染细胞相比，增殖速度更慢，这与Hpr反义mRNA表达及hpr蛋白产生的减少成比例。进行BrdU脉冲处理后，通过流式细胞术分析来估计处于S期的细胞比例。与载体转染细胞相比，Hpr反义转染细胞处于S期的细胞更少。同样在软琼脂中，表达反义cDNA插入片段的细胞形成的集落平均比单独转染载体的细胞少至少7倍。数据表明Hpr抑制剂可能对T细胞白血病具有治疗价值。
