title: Two-dimensional culture of human pancreatic adenocarcinoma cells results in an irreversible transition from epithelial to mesenchymal phenotype.
abstract: Many commercially available cell lines have been in culture for ages, acquiring phenotypes that differ from the original cancers from which these cell lines were derived. Therefore, research on new cell lines could improve the success rates of translational research in cancer. We have developed methods for the isolation and culture of human pancreatic ductal adenocarcinoma (PDAC) cells from murine xenografts of human PDAC. We hypothesize that phenotypes of PDAC cells are modified by in vitro culture conditions over time and by in vivo implantation. Patient-derived xenografts were created in immunodeficient mice using surgically resected tumor specimens. These murine xenografts were then used to establish human PDAC cell lines in culture. Earlier (<5) passage and later (>20) passage cell lines were evaluated separately regarding proliferation, cell cycle, genetic mutations, invasiveness, chemosensitivity, tumorigenesis, epithelial-mesenchymal transition (EMT) status, and proteomics. Later passage cells accelerated their doubling time and colony formation, and were more concentrated in the G0/G1 phase and less in the G2/M checkpoint phase. Later passage cells were more sensitive to gemcitabine and 5-fluorouracil than earlier passage cells, but all four new cell lines were more chemo-resistant compared with commercial ATCC cell lines. EMT induction was observed when establishing and passaging cell lines in vitro and furthermore by growing them as subcutaneous tumors in vivo. This study demonstrates a novel approach to the establishment of PDAC cell lines and observes a process by which newly established cell lines undergo phenotypic changes during in vitro culture and in vivo tumorigenesis. This may help explain differences of treatment effects often observed between experiments conducted in vitro, in vivo, and in human clinical trials.
authors.affiliation: Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Ya'an
authors.initials: Y
authors.lastname: Kang
authors.affiliation: Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Ran
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Department of Gastroenterology and Rheumatology, The Fukushima Medical University School of Medicine, Fukushima, Japan.
authors.forename: Rei
authors.initials: R
authors.lastname: Suzuki
authors.affiliation: Department of Hepatobiliary Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
authors.forename: Shao-qiang
authors.initials: SQ
authors.lastname: Li
authors.affiliation: Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: David
authors.initials: D
authors.lastname: Roife
authors.affiliation: Department of Surgery, Mayo Clinic, Rochester, MN, USA.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Truty
authors.affiliation: Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Deyali
authors.initials: D
authors.lastname: Chatterjee
authors.affiliation: Department of General Surgery, The University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Thomas
authors.affiliation: School of Medicine, Texas Tech Health Sciences Center, Lubbock, TX, USA.
authors.forename: James
authors.initials: J
authors.lastname: Cardwell
authors.affiliation: Neurodiagnostics Lab, The University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Huamin
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Katz
authors.affiliation: Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Jason B
authors.initials: JB
authors.lastname: Fleming
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D003114:Colony-Forming Units Assay; D003841:Deoxycytidine / Q000031:analogs & derivatives; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology*; D005472:Fluorouracil; D064593:Heterografts / Q000166:cytology / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D009361:Neoplasm Invasiveness / Q000503:physiopathology; D010190:Pancreatic Neoplasms / Q000503:physiopathology*; D010641:Phenotype; D040081:Protein Array Analysis; D040901:Proteomics / Q000379:methods; D014407:Tumor Cells, Cultured; D000093542:Gemcitabine
chemical_list: D003841:Deoxycytidine; D005472:Fluorouracil; D000093542:Gemcitabine