FLOW CYTOMETRIC ANALYSIS OF BENIGN AND MALIGNANT TUMORS

在淋巴结阴性的乳腺癌中，有很强的相关性 表现为S期与预后的关系。此外，还展示了一个 非整倍体群体具有诊断意义。我们研究了 二重区分度对S相分数估计的影响 50例乳腺癌四倍体群体的检测 外周血淋巴细胞和细胞系。双倍估计 通过计算机建模和电子检测获得 基于脉冲波形分析的荧光高度VS技术 比较面积和荧光宽度与面积的关系。在生物学上 均质的(如细胞系)样品，所有技术都获得了良好的结果 结果。然而，在生物复杂的样本中，双倍估计 根据方法的不同，差异很大(最多相差五倍) 使用。二重减法的这种变化影响了对 正在增殖的细胞。 轻链限制(检测单抗B细胞)是 在大多数临床情况下是B细胞瘤的代名词。我们 因此，研究了用于轻链检测的流式细胞仪方法 375例患者和181例正常对照。我们在92个细胞中检测到单克隆性 在375例患者样本中，没有正常对照(结果 与临床结果相关)。375份标本中有79份(23%)检出 用标准方法检测的大体单克隆抗体。其中79人 病例中，不同组别的抗轻链试剂有不同 一种检测所有单抗群体的灵敏度，但 与另外两组相比，灵敏度分别为66%和78%。在24年内 标准方法的单克隆性为阴性的情况下， 一名患者有淋巴瘤病史，克隆搜索结果为 根据抗原密度和细胞大小进行。单克隆性是 在这24例中有13例(54%)显示出优越的 该方法的灵敏度。 我们已经建立了流式细胞术方法来定量表达 C-myc、bc1-2和p53蛋白表达的变化。我们将对这些进行比较 方法对分选和未分选的患者标本进行RT-PCR检测。

In lymph node negative breast cancer, a strong association has been shown between S-phase and prognosis. In addition, demonstration of an aneuploid population has diagnostic implications. We studied the effect of doublet discrimination on S-phase fraction estimates and detection of tetraploid populations in fifty breast carcinomas, peripheral blood lymphocytes and cell lines. Doublet estimates obtained using computer modeling as well as electronic detection techniques based upon pulse shape analysis of fluorescent height vs area or fluorescent width vs area were compared. In biologically homogeneous (eg cell lines) specimens, all techniques yielded good results. However, in biologically complex specimens doublet estimates varied greatly (up to five fold difference) depending upon the method used. This variation in doublet subtraction affected estimates of proliferating cells. Light chain restriction (detection of monoclonal B-cells) is synonymous with B-cell neoplasia in most clinical situations. We therefore studied flow cytometric methods for light chain detection in 375 patients and 181 normal controls. We detected monoclonality in 92 of the 375 patient specimens and in no normal controls (results correlated with clinical outcome). 79 out of 375 (23%) specimens were grossly monoclonal and detected by standard methods. Of these 79 cases, different sets of anti-light chain reagents had different sensitivities with one detecting all monoclonal populations but sensitivities of 66% and 78% obtained with two other sets. In 24 cases negative for monoclonality by standard methods but in which the patient had a previous history of lymphoma, a clonal search was performed based upon antigen density and cell size. Monoclonality was detected in 13 of these 24 cases (54%) demonstrating the superior sensitivity of this method. We have established flow cytometric methods to quantitate expression of c-myc, bcl-2 and p53 protein expression. We will compare these methods to RT-PCR methods on sorted and unsorted patient specimens.