Hyakuna N, Naritomi K, Ito E
First Department of Pathology, University of the Ryukyus, School of Medicine, Okinawa, Japan.
Acta Paediatr Jpn. 1998 Aug;40(4):318-23. doi: 10.1111/j.1442-200x.1998.tb01939.x.
Fluorescence in situ hybridization (FISH) has allowed the detection of numerical chromosomal aberrations in interphase nuclei on fresh or frozen smears of leukemia.
To analyze clonality and residual disease in myeloid leukemia retrospectively, we applied FISH to bone marrow smears stored at ambient temperature for up to 9 years.
When hybridization efficiency was investigated on stored control smears from patients without hematological malignancy, more than 96% of nuclei showed the expected number of signals using DNA probes specific for chromosome 7, X or Y. In combination with cell morphology, we observed much higher hybridization efficiency in blasts and granulomonocytic cells compared with lymphoid and erythroid cells. On the basis of good hybridization efficiency for old smear specimens, we applied FISH to stored bone marrow smears of myeloid leukemias, in which either loss of chromosome 7 or loss of sex chromosomes had been verified previously by conventional cytogenetics (one patient with chronic myelomonocytic leukemia (CMML) and four with acute myeloid leukemia (AML; three M2 and one M7)). As a result, the loss of chromosome was detected in blasts from all patients and was observed in mature granulocytes, except in M7. In the CMML patient and one AML (M2) patient with t(8;21), lymphoid and erythroid cells also showed the loss of chromosomes, suggesting that it should occur at stem-cell level. A high amount of residual disease was detected in the morphological remission samples in one AML (M2) patient after induction therapy. The patient eventually succumbed to relapse.
Thus, the present FISH technique is useful to analyze the clinical significance of clonality and the residual disease in myeloid leukemia, retrospectively.
荧光原位杂交（FISH）技术已能够在白血病新鲜或冷冻涂片的间期核中检测染色体数目异常。
为了回顾性分析髓系白血病的克隆性和残留疾病，我们将FISH技术应用于在室温下保存长达9年的骨髓涂片。
在对无血液系统恶性肿瘤患者的对照涂片进行杂交效率研究时，使用针对7号染色体、X或Y染色体的DNA探针，超过96%的细胞核显示出预期的信号数量。结合细胞形态学，我们观察到与淋巴细胞和红细胞相比，原始细胞和粒单核细胞的杂交效率更高。基于旧涂片标本良好的杂交效率，我们将FISH技术应用于髓系白血病保存的骨髓涂片，这些患者先前通过传统细胞遗传学已证实存在7号染色体缺失或性染色体缺失（1例慢性粒单核细胞白血病（CMML）患者和4例急性髓系白血病（AML；3例M2和1例M7）患者）。结果，在所有患者的原始细胞中均检测到染色体缺失，除M7外，在成熟粒细胞中也观察到染色体缺失。在CMML患者和1例伴有t(8;21)的AML（M2）患者中，淋巴细胞和红细胞也显示出染色体缺失，提示这种缺失应发生在干细胞水平。在诱导治疗后，1例AML（M2）患者的形态学缓解样本中检测到大量残留疾病。该患者最终死于复发。
因此，目前的FISH技术可用于回顾性分析髓系白血病克隆性和残留疾病的临床意义。
