Thanee Malinee, Loilome Watcharin, Techasen Anchalee, Namwat Nisana, Boonmars Thidarut, Pairojkul Chawalit, Yongvanit Puangrat
Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand E-mail :
Asian Pac J Cancer Prev. 2015;16(7):3043-50. doi: 10.7314/apjcp.2015.16.7.3043.
The tumor microenvironment (TME) includes numerous non-neoplastic cells such as leukocytes and fibroblasts that surround the neoplasm and influence its growth. Tumor-associated macrophages (TAMs) and cancer-associated fibroblasts (CAFs) are documented as key players in facilitating cancer appearance and progression. Alteration of the macrophage (CD68, CD163) and fibroblast (α-SMA, FSP-1) cells in Opisthorchis viverrini (Ov)-induced cholangiocarcinoma (CCA) was here assessed using liver tissues from an established hamster model and from 43 human cases using immunohistochemistry. We further investigated whether M2-activated TAMs influence CCA cell migration ability by wound healing assay and Western blot analysis. Macrophages and fibroblasts change their phenotypes to M2-TAMs (CD68+, CD163+) and CAFs (α-SMA+, FSP-1+), respectively in the early stages of carcinogenesis. Interestingly, a high density of the M2-TAMs CCA in patients is significantly associated with the presence of extrahepatic metastases (p=0.021). Similarly, CD163+ CCA cells are correlated with metastases (p=0.002), and they may be representative of an epithelial-to-mesenchymal transition (EMT) with increased metastatic activity. We further showed that M2-TAM conditioned medium can induce CCA cell migration as well as increase N-cadherin expression (mesenchymal marker). The present work revealed that significant TME changes occur at an early stage of Ov-induced carcinogenesis and that M2-TAMs are key factors contributing to CCA metastasis, possibly via EMT processes.
肿瘤微环境（TME）包括众多非肿瘤细胞，如围绕肿瘤并影响其生长的白细胞和成纤维细胞。肿瘤相关巨噬细胞（TAMs）和癌症相关成纤维细胞（CAFs）被证明是促进癌症发生和进展的关键因素。本研究使用建立的仓鼠模型的肝脏组织以及43例人类病例的肝脏组织，通过免疫组织化学评估了华支睾吸虫（Ov）诱导的胆管癌（CCA）中巨噬细胞（CD68、CD163）和成纤维细胞（α-SMA、FSP-1）的变化。我们进一步通过伤口愈合试验和蛋白质印迹分析研究了M2激活的TAMs是否影响CCA细胞的迁移能力。在致癌作用的早期阶段，巨噬细胞和成纤维细胞分别将其表型转变为M2-TAMs（CD68+、CD163+）和CAFs（α-SMA+、FSP-1+）。有趣的是，患者中M2-TAMs CCA的高密度与肝外转移的存在显著相关（p=0.021）。同样，CD163+ CCA细胞与转移相关（p=0.002），它们可能代表具有增加转移活性的上皮-间质转化（EMT）。我们进一步表明，M2-TAM条件培养基可以诱导CCA细胞迁移并增加N-钙黏蛋白表达（间充质标志物）。目前的研究表明，在Ov诱导的致癌作用早期会发生显著的TME变化，并且M2-TAMs可能是通过EMT过程促进CCA转移的关键因素。
