INTERPHASE AND METAPHASE PH+ CELLS AND PROGNOSIS

我们研制了一种新的荧光原位杂交探针 （FISH），其从DNA的末端q臂鉴定小于5 Mb的DNA， 人类9号染色体。 9号染色体2/3以上的部分 由该探针鉴定的9号染色体在易位中保留 与CML相关，而不到1/3成为Ph染色体的一部分。 正常人和慢性粒细胞白血病骨髓标本的实验研究 患者们发现这种探针在检测 间期细胞的移位--两个强荧光域 在正常细胞中，两个强域加上第三个弱域， 存在于CML细胞中。 探针的大尺寸还使得能够 高效检测中期细胞中的易位。 在这里，我们将进一步提高探测器的效率， 通过将其与BCR粘粒重叠群在2- 彩色FISH，并通过减少已经很低的不可评分细胞的百分比， 在培养的制剂中。 最重要的是，我们将利用 静止间期Ph+细胞频率测定探针 细胞群以及CML患者中的周期中期细胞， 诊断和治疗后的不同时间点，以确定 哪个参数对预测结果最有效。 患者干部 从与本计划相关的所有不同治疗方案中 将被这样评价。 数据也将与 计划项目，将确定细胞表达的存在 融合蛋白在这些相同的患者组中。 FISH探针也将 用于关联自体骨植入前后Ph+细胞的频率 骨髓移植取得成功。 最有效的鱼 将制作一个用于评估Ph+细胞频率的探针系统 可用于涉及正常细胞与癌细胞分离的项目 或不同谱系或增殖状态的细胞，以及 细胞遗传学核心，用于解决与本计划相关的问题。

We have developed a new probe for fluorescence in situ hybridization (FISH) which identifies less than 5 Mb of DNA from the terminal q-arm of human chromosome 9. More than 2/3 of the portion of chromosome 9 identified by this probe remains with the 9 in the translocation associated with CML while less than 1/3 becomes part of the Ph chromosome. Experiments on bone marrow samples from normal healthy volunteers and CML patients reveal this probe to be highly effective in detecting the translocation in interphase cells -- two intense domains of fluorescence in normal cells while two intense plus a third less intense domain are present in CML cells. The large size of the probe also enables the detection of the translocation in metaphase cells with high efficiency. Here, we will further increase the efficiency of the probe by essentially eliminating false positives by combining it with a BCR cosmid contig in 2- color FISH, and by reducing the already low percentage of unscorable cells in the cultured preparations. Most importantly we will use the ability of the probe to measure the frequencies of Ph+ cells in quiescent interphase populations as well as cycling metaphase cells in CML patients at diagnosis and at various time points after treatment in order to determine which parameter is most effective in predicting outcome. Patient cadres from all the different treatment regimens associated with this Program will be so evaluated. Data will also be compared and evaluated with the Program Project that will be determining the presence of cells expressing fusion protein in these same patient sets. The FISH probe will also be used to relate the frequency of Ph+ cells before and after autologous bone marrow transplantation to successful outcome. The most efficacious FISH probe system for evaluating the frequency of Ph+ cells will be made available to projects involved in separations of normal from cancer cells or cells of different lineages or proliferative states, and to the Cytogenetics Core for their use in problems associated with this Program.