FLOW CYTOMETRIC ANALYSIS OF BENIGN AND MALIGNANT TUMORS

在淋巴结阴性乳腺癌中， 显示在S期和预后之间。 此外，示范一个 非整倍体群体具有诊断意义。 我们研究了 双重鉴别对S相分数估计的影响， 在50个乳腺癌中检测四倍体群体， 外周血淋巴细胞和细胞系。 双重估计 通过计算机建模和电子探测 基于荧光高度与脉冲形状分析的技术 比较面积或荧光宽度与面积。 以生物 均质（如细胞系）标本，所有技术都产生了良好的 结果 然而，在生物学上复杂的样本中， 根据方法的不同，差异很大（高达五倍差异） 采用 双重扣除的这种变化影响了 增殖细胞 轻链限制（检测单克隆B细胞）是 在大多数临床情况下与B细胞瘤形成同义。 我们 因此研究了流式细胞术检测轻链的方法， 375例患者和181例正常对照。 我们在92个样本中检测到单克隆性， 在375例患者标本中，无正常对照（结果 与临床结果相关）。 79/375（23%）例标本 大体上是单克隆的并通过标准方法检测。 其中79 在病例中，不同的抗轻链试剂组具有不同的 灵敏度，一个检测所有单克隆群体，但 另外两组的灵敏度分别为66%和78%。 在24 通过标准方法检测的单克隆性阴性病例，但其中 患者既往有淋巴瘤病史，克隆检索是 基于抗原密度和细胞大小进行。 单克隆性是 在这24例病例中的13例（54%）中检测到，表明上级 这种方法的灵敏度。 我们已经建立了流式细胞术的方法来定量表达 c-myc、bcl-2和p53蛋白表达的变化。 我们将比较这些 方法对已分选和未分选患者标本的RT-PCR方法。

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.