Gaforio José-Juan, Serrano María-José, Sanchez-Rovira Pedro, Sirvent Antonio, Delgado-Rodriguez Miguel, Campos María, de la Torre Nicolás, Algarra Ignacio, Dueñas Rosario, Lozano Ana
Department of Health Sciences, Faculty of Experimental Sciences (Edif. 5), University of Jaén, Campus Las Lagunillas s/n, 23071 Jaén, Spain.
Int J Cancer. 2003 Dec 20;107(6):984-90. doi: 10.1002/ijc.11479.
We investigated whether detection of cytokeratin-positive (CK+) cells in the peripheral blood (PB) of breast cancer patients before chemotherapy could be a prognostic factor. Blood from a total of 92 breast cancer patients was evaluated for the presence of CK+ cells. Blood samples were collected before chemotherapy. Patients entered in the study included: neoadjuvant (n = 25), adjuvant (n = 42) and metastatic (n = 25). Blood samples (10 ml) were centrifuged using a double density-gradient to recovering the mononuclear cell (MNC) and granulocyte cell (GC) fractions. Subsequently, positive immunomagnetic cell separation was carried out to isolating CK+ cells. The enriched cell fraction was cytocentrifuged and then immunocytochemically labeled using an anti-cytokeratin antibody. Our results indicated that breast tumor cells sediment with both MNC and GC fractions. We therefore recommend examination of both fractions in all enrichment protocols. CK+ cells in PB were identified in 57 of 92 (62%) patients when MNC and GC fractions were assessed (range = 1-61 cells, median = 8). No CK+ cells were detected in blood samples of 16 healthy donors. There were significant differences in the presence of CK+ cells according to estrogen receptor expression (p = 0.049), and lymph node status (p = 0.033), but not to the age, menopausal status, type of patient (neoadjuvant, adjuvant or metastatic), TNM stage, histological type, progesterone receptor expression, c-erbB2 expression, p53 expression or Ki67 expression. Regarding the relationship between tumor size (T) and the presence of CK+ cells, a borderline significant trend was observed (p = 0.07). The median follow-up of the patients was 21 months and statistical analysis (Kaplan-Meier analysis) showed that using the method we present, the detection of CK+ cells in PB before starting the chemotherapy in breast cancer patients was significantly correlated with both progression-free survival (p = 0.058) and overall survival (p = 0.003). In conclusion, the present study suggests that detection of CK+ cells in PB before chemotherapy might identify breast cancer patients with poor prognosis.
我们研究了乳腺癌患者化疗前外周血（PB）中细胞角蛋白阳性（CK+）细胞的检测是否可作为一个预后因素。对总共92例乳腺癌患者的血液进行了CK+细胞检测。在化疗前采集血样。纳入研究的患者包括：新辅助治疗（n = 25）、辅助治疗（n = 42）和转移性（n = 25）。将10 ml血样采用双重密度梯度离心以获得单核细胞（MNC）和粒细胞（GC）组分。随后，进行阳性免疫磁珠细胞分选以分离CK+细胞。将富集的细胞组分进行细胞离心涂片，然后用抗细胞角蛋白抗体进行免疫细胞化学标记。我们的结果表明，乳腺肿瘤细胞与MNC和GC组分一起沉降。因此，我们建议在所有富集方案中对这两个组分都进行检测。当评估MNC和GC组分时，92例患者中有57例（62%）在外周血中检测到CK+细胞（范围 = 1 - 61个细胞，中位数 = 8）。在16名健康供者的血样中未检测到CK+细胞。根据雌激素受体表达（p = 0.049）和淋巴结状态（p = 0.033），CK+细胞的存在有显著差异，但与年龄、绝经状态、患者类型（新辅助、辅助或转移性）、TNM分期、组织学类型、孕激素受体表达、c-erbB2表达、p53表达或Ki67表达无关。关于肿瘤大小（T）与CK+细胞存在之间的关系，观察到一种临界显著趋势（p = 0.07）。患者的中位随访时间为21个月，统计分析（Kaplan-Meier分析）表明，采用我们所介绍的方法，乳腺癌患者化疗开始前外周血中CK+细胞的检测与无进展生存期（p = 0.058）和总生存期（p = 0.003）均显著相关。总之，本研究表明化疗前外周血中CK+细胞的检测可能识别出预后不良的乳腺癌患者。
