Usher Suzanne G, Radford Alan D, Villiers Elizabeth J, Blackwood Laura
Small Animal Teaching Hospital, University of Liverpool, The Leahurst Campus, Neston, Wirral, UK.
Exp Hematol. 2009 Jan;37(1):65-77. doi: 10.1016/j.exphem.2008.09.005. Epub 2008 Oct 31.
To determine the frequency of FLT3, C-KIT, and RAS mutations in canine leukemia patients.
Ethylenediamine tetra-acetic acid blood samples were recruited from dogs with suspected leukemia, categorized by quantitative and cytological evaluation and immunophenotyping. Flow cytometry was carried out using antibodies against CD3; CD3e; CD4; CD5; CD8; CD11a, b, c, and d; CD14; CD21; CD34; CD45 and 45RA; CD79a; CD90 (THY-1); major histocompatibility complex II; myeloperoxidase; MAC387; and neutrophil-specific antibody. Genomic DNA was extracted from whole blood and analyzed for mutations in N, H, and K-RAS, FLT3, and C-KIT genes by polymerase chain reaction and sequencing.
Fifty-seven (77.0%) of 74 samples submitted from dogs with suspected leukemia had cytologically and immunophenotypically confirmed leukemia. There were 36 (63.2%) acute leukemias, 16 (28.1%) chronic, 3 (5.3%) prolymphocytic, 1 natural killer cell, and 1 chronic leukemia undergoing blast transformation. N-RAS mis-sense mutations were identified in 14 (25%) dogs with acute myeloid (AML) or lymphoid (ALL) leukemia, and also in one dog in the leukemic phase of lymphoma. Mutations in K-RAS were found in two dogs with AML. There were no H-RAS mutations. FLT3 internal tandem duplications were identified in three dogs with ALL, and a mis-sense mutation was found in one dog with ALL. C-KIT mutations were identified in three dogs with AML. Sixty-one percent of dogs with acute leukemia harbored mutations in N/K-RAS, FLT3, or C-KIT.
RAS, FLT3, and C-KIT mutations, analogous to those found in human leukemia, occur commonly in acute canine leukemia.
确定犬白血病患者中FLT3、C-KIT和RAS基因突变的频率。
从疑似白血病犬采集乙二胺四乙酸血样，通过定量、细胞学评估和免疫表型分析进行分类。使用抗CD3、CD3e、CD4、CD5、CD8、CD11a、b、c和d、CD14、CD21、CD34、CD45和45RA、CD79a、CD90（THY-1）、主要组织相容性复合体II、髓过氧化物酶、MAC387和中性粒细胞特异性抗体进行流式细胞术检测。从全血中提取基因组DNA，通过聚合酶链反应和测序分析N、H和K-RAS、FLT3和C-KIT基因的突变情况。
74份来自疑似白血病犬的样本中，57份（77.0%）经细胞学和免疫表型分析确诊为白血病。其中急性白血病36份（63.2%），慢性白血病16份（28.1%），原淋巴细胞白血病3份（5.3%），自然杀伤细胞白血病1份，慢性白血病急变1份。在14只患有急性髓性白血病（AML）或淋巴细胞白血病（ALL）的犬中以及1只处于淋巴瘤白血病期的犬中发现了N-RAS错义突变。在2只患有AML的犬中发现了K-RAS突变。未发现H-RAS突变。在3只患有ALL的犬中发现了FLT3内部串联重复，在1只患有ALL的犬中发现了错义突变。在3只患有AML的犬中发现了C-KIT突变。61%的急性白血病犬在N/K-RAS、FLT3或C-KIT基因中存在突变。
与人类白血病中发现的类似，RAS、FLT3和C-KIT基因突变在急性犬白血病中普遍存在。
