Brotherick I, Browell D A, Shenton B K, Egan M, Cunliffe W J, Webb L A, Lunt L G, Young J R, Higgs M J
Department of Surgery, University of Newcastle upon Tyne, UK.
Br J Cancer. 1998 May;77(10):1657-60. doi: 10.1038/bjc.1998.272.
The effect of 3-week, preoperative tamoxifen treatment on oestrogen receptor (ER) levels, expressed by primary breast tumours, was examined. Patients (age-matched) with breast cancer, confirmed by fine-needle aspiration, were either treated with 20 mg ml(-1) oral tamoxifen per day or received no medication in the 3-week interval between assessment and surgery. Quantification of ER using flow cytometry was performed on the surgically removed tumour samples from tamoxifen-treated (n = 40) and control (n = 38, untreated) patient groups. The tumours were mechanically disaggregated, and saponin treatment rendered these cells permeable to antibodies. Using dual-parameter labelling with a FITC-conjugated antibody (NCL-5D3) directed against cytokeratin 8/18/19 and a biotinylated antibody (DAKO-ER 1D5) directed against the oestrogen receptor, ER quantification was determined on a number of receptors per cell basis. Using QC quantum bead standards, ER levels in the epithelial cell population, the non-epithelial cell population and the whole-cell population (ER+) were calculated. ER levels were significantly lower in the total cell population than tamoxifen-treated patients (P = 0.002) when compared with the control (untreated) group. By using a gating procedure using 5D3 antibody positivity, a significantly lower level was detected on examining the cytokeratin-positive population alone (P = 0.006). Using a complementary gating technique, ER levels were quantified in the cytokeratin-negative cell population. Examination of this group of cells showed no significant difference between the levels of oestrogen receptor found in the tamoxifen-treated and untreated groups (P = 0.4). We have demonstrated that ER levels can be monitored by flow cytometry. ER levels in patients treated with tamoxifen 3 weeks before operation are significantly lower than in a comparative group of patients who received no drug. Furthermore, the most significant difference in receptor levels is seen by quantification of total ER levels expressed by all the tissue.
研究了术前3周他莫昔芬治疗对原发性乳腺肿瘤所表达的雌激素受体（ER）水平的影响。经细针穿刺确诊为乳腺癌的患者（年龄匹配），在评估与手术之间的3周间隔期内，每天接受20 mg/ml口服他莫昔芬治疗，或不接受任何药物治疗。对来自他莫昔芬治疗组（n = 40）和对照组（n = 38，未治疗）患者的手术切除肿瘤样本，使用流式细胞术对ER进行定量分析。将肿瘤机械分散，皂角苷处理使这些细胞对抗体具有通透性。使用针对细胞角蛋白8/18/19的异硫氰酸荧光素（FITC）偶联抗体（NCL-5D3）和针对雌激素受体的生物素化抗体（DAKO-ER 1D5）进行双参数标记，以每个细胞上的受体数量为基础确定ER定量。使用QC量子微珠标准，计算上皮细胞群体、非上皮细胞群体和全细胞群体（ER+）中的ER水平。与对照组（未治疗）相比，总细胞群体中的ER水平显著低于他莫昔芬治疗的患者（P = 0.002）。通过使用基于5D3抗体阳性的门控程序，单独检查细胞角蛋白阳性群体时检测到显著较低的水平（P = 0.006）。使用互补的门控技术，对细胞角蛋白阴性细胞群体中的ER水平进行定量。对这组细胞的检查显示，他莫昔芬治疗组和未治疗组中发现的雌激素受体水平无显著差异（P = 0.4）。我们已经证明，可以通过流式细胞术监测ER水平。术前3周接受他莫昔芬治疗的患者的ER水平显著低于未接受药物治疗的对照组患者。此外，通过对所有组织所表达的总ER水平进行定量分析，可以观察到受体水平的最显著差异。
