Davidson Ben, Konstantinovsky Sophya, Nielsen Søren, Dong Hiep Phuc, Berner Aasmund, Vyberg Mogens, Reich Reuven
Department of Pathology, The Norwegian Radium Hospital, Montebello, University of Oslo, Oslo, Norway.
Clin Cancer Res. 2004 Nov 1;10(21):7335-46. doi: 10.1158/1078-0432.CCR-04-0183.
The aim of this study was to characterize phenotypic alterations along the progression of breast carcinoma from primary tumor to pleural effusion through analysis of the expression of proteases, laminin receptors (LRs), and transcription factors involved in invasion and metastasis.
The material studied consisted of 60 malignant pleural effusions from breast cancer patients and 68 corresponding solid tumors (37 primary and 31 metastatic tumors). Expression of matrix metalloproteinases [MMPs (MMP-1, MMP-2, MMP-9, and MMP-14)], the MMP inhibitor tissue inhibitor of metalloproteinase-2, the MMP inducer EMMPRIN, the 67-kDa LR, the alpha6 integrin subunit, and the transcription factors AP-2, Ets-1, and PEA3 was studied using immunohistochemistry, mRNA in situ hybridization, reverse transcription-polymerase chain reaction, zymography, and flow cytometry. Hormone receptor (estrogen receptor and progesterone receptor) status and c-erbB-2 status were also studied.
Significantly reduced estrogen receptor (P < 0.001) and progesterone receptor (P = 0.001) expression was seen in effusions compared with primary tumors, with opposite findings for c-erbB-2 (P = 0.003). Tumor cell MMP-2 protein expression in effusions was higher than that in primary tumors (P < 0.001) and lymph node metastases (P = 0.01). In situ hybridization demonstrated higher MMP-2 (P = 0.007), PEA3 (P = 0.038), and EMMPRIN (P = 0.026) mRNA expression in effusions. The time to progression from primary tumor to effusion was significantly shorter for patients whose primary tumors expressed MMP-1 (P = 0.016) and who expressed the 67-kDa LR protein in primary tumor (P = 0.007) and effusion (P = 0.015).
Our data provide documented evidence of molecular events that occur during the progression of breast carcinoma from primary tumor to effusion. The coordinated up-regulation of MMP-2 and Ets transcription factors in carcinoma cells in effusions is in full agreement with our previous reports linking these factors to poor prognosis in ovarian cancer. The rapid progression to effusion in cases showing MMP-1 and 67-kDa LR expression in primary tumor cells links aggressive clinical behavior with expression of metastasis-associated molecules in this setting.
本研究旨在通过分析参与侵袭和转移的蛋白酶、层粘连蛋白受体（LRs）及转录因子的表达，来描述乳腺癌从原发性肿瘤进展至胸腔积液过程中的表型改变。
所研究的材料包括60例乳腺癌患者的恶性胸腔积液以及68例相应的实体瘤（37例原发性肿瘤和31例转移性肿瘤）。采用免疫组织化学、mRNA原位杂交、逆转录-聚合酶链反应、酶谱分析及流式细胞术，研究基质金属蛋白酶[MMPs（MMP-1、MMP-2、MMP-9和MMP-14）]、MMP抑制剂金属蛋白酶组织抑制剂-2、MMP诱导剂EMMPRIN、67-kDa LR、α6整合素亚基以及转录因子AP-2、Ets-1和PEA3的表达。同时也研究了激素受体（雌激素受体和孕激素受体）状态及c-erbB-2状态。
与原发性肿瘤相比，胸腔积液中雌激素受体（P < 0.001）和孕激素受体（P = 0.001）表达显著降低，而c-erbB-2情况则相反（P = 0.003）。胸腔积液中肿瘤细胞MMP-2蛋白表达高于原发性肿瘤（P < 0.001）和淋巴结转移灶（P = 0.01）。原位杂交显示胸腔积液中MMP-2（P = 0.007）、PEA3（P = 0.038）和EMMPRIN（P = 0.026）mRNA表达更高。原发性肿瘤表达MMP-1（P = 0.016）以及原发性肿瘤（P = 0.007）和胸腔积液（P = 0.015）中表达67-kDa LR蛋白的患者，从原发性肿瘤进展至胸腔积液的时间显著缩短。
我们的数据提供了乳腺癌从原发性肿瘤进展至胸腔积液过程中发生的分子事件的文献证据。胸腔积液中癌细胞内MMP-2和Ets转录因子的协同上调与我们之前将这些因子与卵巢癌不良预后相关联的报道完全一致。原发性肿瘤细胞中显示MMP-1和67-kDa LR表达的病例迅速进展至胸腔积液，表明在这种情况下侵袭性临床行为与转移相关分子的表达有关。
