Institute of Anatomy, First Faculty of Medicine, Charles University, Prague, Czech Republic.
Department of Otorhinolaryngology, Head and Neck Surgery, First Faculty of Medicine, Charles University and University Hospital Motol, Prague, Czech Republic.
Cancer Genomics Proteomics. 2021 May-Jun;18(3):221-243. doi: 10.21873/cgp.20254.
BACKGROUND/AIM: Pancreatic ductal adenocarcinoma (PDAC) still represents one of the most aggressive cancers. Understanding of the epithelial-mesenchymal crosstalk as a crucial part of the tumor microenvironment should pave the way for therapies to improve patient survival rates. Well-established cell lines present a useful and reproducible model to study PDAC biology. However, the tumor-stromal interactions between cancer cells and cancer-associated fibroblasts (CAFs) are still poorly understood.
We studied interactions between four PDAC cell lines (Panc-1, CAPAN-2, MIAPaCa-2, and PaTu-8902) and conditioned media derived from primary cultures of normal fibroblasts/PDAC-derived CAFs (PANFs).
When the tested PDAC cell lines were stimulated by PANF-derived conditioned media, the most aggressive behavior was acquired by the Panc-1 cell line (increased number and size of colonies, remaining expression of vimentin and keratin 8 as well as increase of epithelial-to-mesenchymal polarization markers), whereas PaTu-8902 cells were rather inhibited. Of note, administration of the conditioned media to MIAPaCa-2 cells resulted in an inverse effect on the size and number of colonies, whereas CAPAN-2 cells were rather stimulated. To explain the heterogeneous pattern of the observed PDAC crosstalk at the in vitro level, we further compared the phenotype of primary cultures of cells derived from ascitic fluid with that of the tested PDAC cell lines, analyzed tumor samples of PDAC patients, and performed gene expression profiling of PANFs. Immuno-cyto/histo-chemical analysis found specific phenotype differences within the group of examined patients and tested PDAC cell lines, whereas the genomic approach in PANFs found the key molecules (IL6, IL8, MFGE8 and periostin) that may contribute to the cancer aggressive behavior.
The desmoplastic patient-specific regulation of cancer cells by CAFs (also demonstrated by the heterogeneous response of PDAC cell lines to fibroblasts) precludes simple targeting and development of an effective treatment strategy and rather requires establishment of an individualized tumor-specific treatment protocol.
背景/目的：胰腺导管腺癌（PDAC）仍然是最具侵袭性的癌症之一。对上皮-间充质串扰的理解作为肿瘤微环境的重要组成部分，应该为提高患者生存率的治疗方法铺平道路。成熟的细胞系是研究 PDAC 生物学的有用且可重复的模型。然而，癌细胞与癌相关成纤维细胞（CAF）之间的肿瘤-基质相互作用仍知之甚少。
我们研究了四种 PDAC 细胞系（Panc-1、CAPAN-2、MIAPaCa-2 和 PaTu-8902）与源自正常成纤维细胞/PDAC 衍生 CAF（PANF）的原代培养物的条件培养基之间的相互作用。
当测试的 PDAC 细胞系被 PANF 衍生的条件培养基刺激时，Panc-1 细胞系获得了最具侵袭性的行为（增加了集落的数量和大小，仍然表达波形蛋白和角蛋白 8 以及增加上皮-间充质极化标记物），而 PaTu-8902 细胞则受到抑制。值得注意的是，将条件培养基施用于 MIAPaCa-2 细胞导致集落的大小和数量出现相反的效果，而 CAPAN-2 细胞则受到刺激。为了解释在体外水平观察到的 PDAC 串扰的异质模式，我们进一步比较了来自腹水的细胞原代培养物的表型与测试的 PDAC 细胞系的表型，分析了 PDAC 患者的肿瘤样本，并对 PANFs 进行了基因表达谱分析。免疫细胞化学/组织化学分析在检查的患者和测试的 PDAC 细胞系组内发现了特定的表型差异，而 PANFs 的基因组方法发现了可能导致癌症侵袭性行为的关键分子（IL6、IL8、MFGE8 和 periostin）。
CAF 对癌细胞的促纤维化患者特异性调节（也通过 PDAC 细胞系对成纤维细胞的异质反应证明）排除了简单的靶向和有效治疗策略的开发，而是需要建立个体化的肿瘤特异性治疗方案。
