Cierna Zuzana, Mego Michal, Janega Pavol, Karaba Marian, Minarik Gabriel, Benca Juraj, Sedlácková Tatiana, Cingelova Silvia, Gronesova Paulina, Manasova Denisa, Pindak Daniel, Sufliarsky Jozef, Danihel Ludovit, Reuben James M, Mardiak Jozef
Translational Research Unit, Comenius University, Bratislava, Slovakia.
BMC Cancer. 2014 Jun 28;14:472. doi: 10.1186/1471-2407-14-472.
Matrix metalloproteinases (MMPs) are involved in cancer invasion and metastasis. Circulating tumor cells (CTCs) play role in tumor dissemination and are an independent survival predictor in breast cancer (BC) patients. The aim of this study was to assess correlation between CTCs and tumor MMP1 in BC.
Study included 149 primary BC patients treated by surgery from March 2012 to March 2013. Peripheral blood mononuclear cells (PBMC) were depleted of hematopoietic cells using RossetteSep(TM) selection kit. RNA extracted from CD45-depleted PBMC was interrogated for expression of EMT (TWIST1, SNAIL1, SLUG, ZEB1) and epithelial (CK19) gene transcripts by qRT-PCR. Patient samples with higher epithelial and/or mesenchymal gene transcripts than those of healthy donors (n = 60) were considered as CTC positive. Expression of MMP1 in surgical specimens was evaluated by immunohistochemistry.
CTCs were detected in 24.2% patients. CTCs exhibiting only epithelial markers were present in 8.7% patients, whereas CTCs with epithelial-mesenchymal transition (EMT) markers (CTC_EMT) were observed in 13.4% of patients and CTCs co-expressing both markers were detected in 2.0% patients. Patients with CTC_EMT in peripheral blood had significantly increased expression of MMP1 in tumor cells (p = 0.02) and tumor associated stroma (p = 0.05) than those of patients without CTC_EMT. In multivariate analysis, CTC_EMT and tumor grade were independently associated with MMP1 expression in cancer cells, while CTC_EMT and Ki67 were independently associated with MMP1 expression in cancer associated stroma.
Our data suggest link between MMP1 and CTCs with EMT phenotype and support role of MMPs and EMT in tumor dissemination.
基质金属蛋白酶（MMPs）参与癌症侵袭和转移。循环肿瘤细胞（CTCs）在肿瘤播散中起作用，并且是乳腺癌（BC）患者独立的生存预测指标。本研究旨在评估BC患者中CTCs与肿瘤MMP1之间的相关性。
研究纳入了2012年3月至2013年3月期间接受手术治疗的149例原发性BC患者。使用罗斯特分离（RossetteSep™）选择试剂盒去除外周血单个核细胞（PBMC）中的造血细胞。通过qRT-PCR检测从去除CD45的PBMC中提取的RNA中EMT（TWIST1、SNAIL1、SLUG、ZEB1）和上皮（CK19）基因转录本的表达。上皮和/或间充质基因转录本高于健康供体（n = 60）的患者样本被视为CTCs阳性。通过免疫组织化学评估手术标本中MMP1的表达。
24.2%的患者检测到CTCs。仅表现上皮标志物的CTCs存在于8.7%的患者中，而具有上皮-间充质转化（EMT）标志物（CTC_EMT）的CTCs在13.4%的患者中观察到，同时表达两种标志物的CTCs在2.0%的患者中检测到。外周血中具有CTC_EMT的患者肿瘤细胞中MMP1的表达（p = 0.02）和肿瘤相关基质中MMP1的表达（p = 0.05）显著高于无CTC_EMT的患者。在多变量分析中，CTC_EMT和肿瘤分级与癌细胞中MMP1的表达独立相关，而CTC_EMT和Ki67与癌相关基质中MMP1的表达独立相关。
我们的数据表明MMP1与具有EMT表型的CTCs之间存在联系，并支持MMPs和EMT在肿瘤播散中的作用。
