O'Brien M C, Gupta R K, Lee S Y, Bolton W E
Coulter Corporation, Immunology Research and Technology, Miami, Florida, USA.
Cytometry. 1995 Sep 1;21(1):76-83. doi: 10.1002/cyto.990210115.
The exclusion of non-tumor and dead cells from the analysis of live tumor cells can significantly improve the accuracy of prognostic indicators such as proliferative and DNA indexes. To target live breast tumor cells in a heterogeneous breast tumor model, we have designed a panel consisting of the DNA-specific dye DAPI and epithelial tissue-specific (cytokeratin), tumor-associated (MC5), proliferation-associated (proliferating cell nuclear antigen), and viability-associated (tubulin) markers. The breast tumor model consisted of a mixture of equal numbers of live and dead MDA-MB-175-VII (breast tumor) cells, live CEM (leukemic) cells, and live peripheral blood mononuclear cells. Targeting the live MDA cells in the mixture by gating on tubulin, cytokeratin, and MC5 resulted in a sevenfold increase in PCNA positivity (from 3% ungated to 22.3%), a 60% decrease in the %S-phase fraction (from 37.2% ungated to 15%), and elimination of extraneous hypodiploid and diploid components, enriching the tetraploid MDAs. These results are consistent with those obtained from unmixed live MDA cells. The combined utilization of this panel and "cumulative" electronic gating of the targeted population increases the number of relevant parameters that can be analyzed per sample and the accuracy of the resultant data.
在对活肿瘤细胞的分析中排除非肿瘤细胞和死亡细胞，可显著提高诸如增殖指数和DNA指数等预后指标的准确性。为了在异质性乳腺肿瘤模型中靶向活乳腺肿瘤细胞，我们设计了一个组合，其中包括DNA特异性染料DAPI以及上皮组织特异性（细胞角蛋白）、肿瘤相关（MC5）、增殖相关（增殖细胞核抗原）和活力相关（微管蛋白）标志物。乳腺肿瘤模型由等量的活的和死亡的MDA-MB-175-VII（乳腺肿瘤）细胞、活的CEM（白血病）细胞和活的外周血单个核细胞混合而成。通过根据微管蛋白、细胞角蛋白和MC5进行门控来靶向混合物中的活MDA细胞，导致PCNA阳性率增加了7倍（从非门控时的3%增至22.3%），S期分数降低了60%（从非门控时的37.2%降至15%），并消除了无关的亚二倍体和二倍体成分，富集了四倍体MDA细胞。这些结果与从未混合的活MDA细胞获得的结果一致。该组合与靶向群体的“累积”电子门控的联合使用增加了每个样本可分析的相关参数数量以及所得数据的准确性。
