Kimmig R, Spelsberg H, Kapsner T, Untch M, Hepp H
Department of Obstetrics and Gynecology, Klinikum Grosshadern, Ludwig-Maximilians-University, Munich, Germany.
Anal Cell Pathol. 1994 Oct;7(3):205-15.
Flow cytometric assessment of DNA-ploidy and S-phase fraction in breast cancer is compromised by the heterogeneity of cell subpopulations derived from the malignant and surrounding connective tissue, e.g. tumour, stromal and inflammatory cells. To identify tumour cell subpopulations, epithelial cells were labeled by a FITC-conjugated cytokeratin antibody (CK6, CK18) prior to flow cytometric cell cycle analysis in 205 fresh specimens of primary breast cancer. We found 158/205 (77%) DNA-aneuploid tumours compared to 127/205 (62%) without identification of cytokeratin positive cells (P < 0.001). In addition, the number of detected DNA-multiploid tumours rose from 31 (15%) to 51 (25%) after gating for cytokeratin positive cells. In DNA-diploid tumours, S-phase and G2M-phase fractions were significantly higher in cytokeratin positive (tumour) cells compared to total cell populations (4.4% and 5.8% vs. 3.2% and 4.4%; P < 0.001). Cytokeratin negative cells were found in all tumours and can be used as internal standard for calculation of ploidy and for quality control (CV, linearity) of each individual sample. We conclude that at least 20% of DNA-aneuploid tumours would not have been diagnosed without cytokeratin labeling. In addition, influence of non-tumourous cell elements on cell cycle analysis can be markedly reduced. Therefore, both determination of DNA-ploidy and cell cycle analysis can be optimized by cytokeratin labeling.
乳腺癌中DNA倍性和S期分数的流式细胞术评估受到源自恶性和周围结缔组织的细胞亚群（如肿瘤细胞、基质细胞和炎性细胞）异质性的影响。为了识别肿瘤细胞亚群，在对205例原发性乳腺癌新鲜标本进行流式细胞术细胞周期分析之前，用异硫氰酸荧光素（FITC）偶联的细胞角蛋白抗体（CK6、CK18）标记上皮细胞。我们发现，与127/205例（62%）未鉴定出细胞角蛋白阳性细胞的肿瘤相比，158/205例（77%）为DNA非整倍体肿瘤（P<0.001）。此外，在选通细胞角蛋白阳性细胞后，检测到的DNA多倍体肿瘤数量从31例（15%）增加到51例（25%）。在DNA二倍体肿瘤中，细胞角蛋白阳性（肿瘤）细胞的S期和G2M期分数显著高于总细胞群（分别为4.4%和5.8%，而总细胞群为3.2%和4.4%；P<0.001）。在所有肿瘤中均发现了细胞角蛋白阴性细胞，其可作为计算倍性的内标以及每个样本的质量控制（变异系数、线性）指标。我们得出结论，若不进行细胞角蛋白标记，至少20%的DNA非整倍体肿瘤将无法被诊断出来。此外，非肿瘤细胞成分对细胞周期分析的影响可显著降低。因此，通过细胞角蛋白标记可优化DNA倍性测定和细胞周期分析。
