CORE--HYPERMETAPHASE FISH AND PCR IN ACUTE MYELOGENOUS LEUKEMIA

荧光原位杂交(FISH)方法将进一步 开发和利用作为一项主要程序的高 急性髓系白血病细胞周期频率的分辨率量化。急性髓细胞白血病 与下列染色体异常相关的疾病 将通过此程序进行研究-inv(16)，t8；21，t15；17，+8， 和-7。这一过程是基于我们在CML中的结果，我们有 结合长期有丝分裂抑制产生了数千个 使用FISH探针的中期/幻灯片(“超中期”) 检测与基因重组相关的染色体重排 恶性(超中期/FISH或HMF)。在CML模型中，我们有 证明癌细胞可以很容易地在 循环病人骨髓标本中的细胞。这个 程序将允许检测1%的癌症、电话和遗嘱 允许监测此类单元格频率的4%变化。 该方法将在这里应用，以确定 急性髓系白血病患者在确诊时的疾病和评估 急性髓系白血病患者治疗过程中的治疗效果。这个 来自+8和-7的数据将与间期FISH相关联 由Core C生成的相同样本的数据。此外，我们 将进行逆转录，然后进行聚合酶链式反应(RT/PCR)以 测定T15；17和T8；21的嵌合转录本水平 AML与HMF数据以及间期FISH相关 和标准G-带细胞遗传学(CG)数据在同一 最终我们将完成开发和进行 对inv(16)样本进行基因组聚合酶链式反应以评估细胞水平 也包含对我们的HMF结果有反应的嵌合基因 对同一样本进行RT/PCR、间期FISH和CG检测。 实验和相关性将用 项目1和项目2确定的患者的进展(Estey博士 和Champlin分别)，以确定收益和 每种技术在建立 患者对所涉及治疗的细胞遗传学反应。这些 程序将给我们新的见解来确定 临床有意义的残留病水平，并指导 急性髓系白血病患者个体恶性肿瘤的进一步处理。

A fluorescence in-situ hybridization (FISH) method will be further developed and utilized as a major procedure for the high resolution quantitation of the frequency of cycling AML cells. AML diseases associated with the following chromosomal abnormalities will be studied by this procedure - inv(16), t8;21, t15;17, +8, and -7. The procedure is based on our results in CML where we have coupled long term mitotic arrest to produce thousands of metaphases/slide ("hypermetaphase") with the use of a FISH probe that detects the chromosomal rearrangement associated with the malignancy (hypermetaphase/FISH or HMF). In the CML model we have demonstrated that cancer cells could be readily quantitated in cycling cells from patient bone marrow preparations. The procedures will allow detection of <1% cancer, calls and will permit monitoring of <4% changes in the frequency of such cells. The methodology will be applied here to identify severity of disease in AML patients at diagnosis and to evaluate the effectiveness of therapies on AML patients during treatment. The data from +8 and -7 will be correlated with the interphase FISH data on those same samples generated by Core C. In addition we will conduct reverse transcription followed by PCR (RT/PCR) to determine the level of chimeric transcript in t15;17 and t8;21 AMLs to correlate with the HMF data as well as the interphase FISH and standard G-band cytogenetics (CG) data generated on the same samples by Core Finally we will complete development and conduct genomic PCR on inv(16) samples to evaluate the level of cells containing the chimeric gene reactive to our HMF results as well as RT/PCR, interphase FISH and CG conducted on those same samples. The experiments and correlations will be evaluated with the progress of the patients determined by Projects 1 and 2 (Drs.Estey and Champlin respectively) to determine the benefits and shortcomings of each of the technologies in establishing the cytogenetic response of patients to therapies involved. These procedures will give us new insights into determining the clinically significant level of residual disease, and guide the further management of individual malignancies in AML patients.