Kalmanti M, Kalmantis T, Vassilaki M, Galanopoulos A, Grenzelias D, Spandidos D A
Department of Pediatric Hematology-Oncology, University of Crete, Medical School, Heraklion, Greece.
Anticancer Res. 1992 Nov-Dec;12(6B):2177-80.
Point mutations of the ras genes have been detected in various hematologic malignancies. This genetic event may either occur in all malignant cells or be acquired by different subclones, which however, cannot be demonstrated adequately by analyzing only DNA derived from patient specimens. The availability of the ras p21 monoclonal antibody (MoAb) Y 13259 makes possible the direct study of the distribution of the ras gene product in human malignant cells. In this report the expression of the ras p21 oncoprotein in the bone marrow smears of 35 children with acute leukemia has been analyzed. The smears were treated with the MoAb Y 13259, biotinylated goat anti-rat IgG, streptavidin, peroxidase and stained with diaminobenzidine (DAB). The intensity of the staining was evaluated by two independent observers as negative or equivocal (-/+), moderate (+) or intense (++), by counting one thousand cells. Patients were also classified according to the percentage of the stained cells into four groups (0, I, II, III). It was found that 22/35 (63%) were (+) or (++) positive as follows: 11/21 (52%) with ALL CALLA (+), 2/2 ALL-B, 3/3 ALL-T and 6/9 AML. In Group 0 (none of the blasts was stained) were 13/35 (37%), as well as in Group I (1 to 25% of the blasts stained 1+ or 2+ positive), while in Group II (26 to 50% positive stained) 3/35 and in Group III (more than 51% stained) 6/35, all of which were AML (6/9). It is concluded that the immunohistochemical analysis of the ras p21 in blast cells of children with acute leukemia may demonstrate that ras gene expression in some subclones, the intensity and percentage of which may be of some clinical importance.
在各种血液系统恶性肿瘤中均检测到了ras基因的点突变。这一基因事件可能发生于所有恶性细胞中，也可能由不同的亚克隆获得，然而，仅通过分析患者标本来源的DNA并不能充分证明这一点。ras p21单克隆抗体（MoAb）Y 13259的可用性使得直接研究ras基因产物在人类恶性细胞中的分布成为可能。在本报告中，分析了35例急性白血病患儿骨髓涂片上ras p21癌蛋白的表达情况。涂片用MoAb Y 13259、生物素化山羊抗大鼠IgG、链霉亲和素、过氧化物酶处理，并用二氨基联苯胺（DAB）染色。两名独立观察者通过计数1000个细胞，将染色强度评估为阴性或可疑（-/+）、中度（+）或强阳性（++）。患者还根据染色细胞的百分比分为四组（0、I、II、III）。结果发现，22/35（63%）为（+）或（++）阳性，具体如下：11/21（52%）为ALL CALLA（+）、2/2为ALL-B、3/3为ALL-T以及6/9为AML。0组（无原始细胞染色）为13/35（37%），I组（1%至25%的原始细胞染色为1+或2+阳性）也如此，而II组（26%至50%阳性染色）为3/35，III组（超过51%染色）为6/35，所有这些均为AML（6/9）。结论是，对急性白血病患儿原始细胞中ras p21进行免疫组化分析可能表明，ras基因在某些亚克隆中的表达，其强度和百分比可能具有一定临床意义。
