Lubiński J, Chosia M, Kotańska K, Huebner K
Department of Tumor Pathology, Medical Academy, Szczecin, Poland.
Anal Quant Cytol Histol. 1988 Dec;10(6):383-90.
DNA was isolated from 20 fine needle aspiration (FNA) biopsies from lymphomas, hyperplastic lymph nodes and nonlymphoid malignant tumors. Small aliquots (0.2 microgram to 2.0 micrograms) of DNA from each sample were digested to completion with restriction endonuclease Eco RI and/or Bam HI and electrophoresed in 0.8% agarose minigels. DNA was transferred to a nylon filter after brief treatment in HCl and subsequent denaturation and neutralization. Filters were hybridized to radiolabeled JH, C kappa, TCR beta or bcl-2 probes to determine if these genes were in germline or rearranged configurations in each of the samples. It was possible to demonstrate rearrangement of at least one immunoglobulin gene in each of the samples diagnosed as lymphoma, while all samples derived from hyperplastic lymph nodes and nonlymphoid malignant tumors exhibited a germline pattern for each probe tested. Thus, FNA biopsies can provide suitable and sufficient DNA for genotypic analysis using molecular probes that detect gene rearrangement.
从淋巴瘤、增生性淋巴结和非淋巴恶性肿瘤的20份细针穿刺(FNA)活检样本中分离出DNA。每个样本的小份DNA(0.2微克至2.0微克)用限制性内切酶Eco RI和/或Bam HI完全消化,并在0.8%琼脂糖微型凝胶中进行电泳。DNA在HCl中短暂处理、随后变性和中和后转移到尼龙滤膜上。滤膜与放射性标记的JH、C kappa、TCR beta或bcl-2探针杂交,以确定这些基因在每个样本中是处于种系构型还是重排构型。在每个诊断为淋巴瘤的样本中都有可能证明至少一个免疫球蛋白基因发生了重排,而所有来自增生性淋巴结和非淋巴恶性肿瘤的样本对每个测试探针都呈现种系模式。因此,FNA活检可以为使用检测基因重排的分子探针进行基因分型分析提供合适且足够的DNA。