Abstract
Pancreatic ductal adenocarcinoma (PDAC) is characterized by an extremely poor prognosis, and its treatment remains a challenge. As the existing in vitro experimental models offer only a limited resemblance to human PDAC, there is a strong need for additional research tools to better understand PDAC tumor biology, particularly the impact of the tumor stroma. Here, we report for the first time the establishment and characterization of human PDAC-derived paired primary monolayer cultures of (epithelial) cancer cells (PCCs) and mesenchymal stellate cells (PSCs) derived from the same tumor by the outgrowth method. Characterization of cell morphology, cytostructural, and functional profiles and proteomics-based secretome analysis were performed. All PCCs harbored KRAS and TP53 mutations, and expressed cytokeratin 19, ki-67, and p53, while the expression of EpCAM and vimentin was variable. All PSCs expressed α-smooth muscle actin (α-SMA) and vimentin. PCCs showed a significantly higher growth rate and proliferation than PSCs. Secretome analysis confirmed the distinct nature of PCCs as compared to PSCs and allowed identification of potential molecular regulators of PSC-conditioned medium (PSC-CM)-induced migration of PCCs. Paired primary cultures of PCCs and PSCs derived from the same tumor specimen represent a novel experimental model for basic research in PDAC tumor biology.
Highlights
Pancreatic ductal adenocarcinoma (PDAC), commonly known as pancreatic cancer, is one of the most lethal solid tumors, characterized by early metastasis, a complex tumor microenvironment, profound chemoresistance, and overall 5-year survival of less than 7% [1,2]
Reagents were purchased from the following sources: Dulbecco’s modified Eagle’s medium (DMEM) containing 4.5 g/L glucose, penicillin-streptomycin (Pen-Strep), Amphotericin B, Trypsin/EDTA, fetal bovine serum (FBS), and PierceTM BCA protein assay kit from Thermo Fisher Scientific (Waltham, MA, USA); bovine serum albumin (BSA), 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), and phosphate-buffered saline (PBS) from Sigma-Aldrich (St Louis, MO, USA); and Ultima Gold from Perkin Elmer (Waltham, MA, USA)
The study of the cellular interactions taking place between the cancer and stroma has recently emerged as an important topic in pancreatic cancer research [43,44,45]
Summary
Pancreatic ductal adenocarcinoma (PDAC), commonly known as pancreatic cancer, is one of the most lethal solid tumors, characterized by early metastasis, a complex tumor microenvironment, profound chemoresistance, and overall 5-year survival of less than 7% [1,2]. Despite new insight in the genomic basis of the disease, therapeutic advancement for PDAC has been negligible over the past decades [3,4,5]. PDAC is the fourth most common cause of cancer deaths in the Western world and is expected to rank second by 2030 [6]. The profound resistance of PDAC to conventional chemotherapy is considered a major impediment to improved survival [10]
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