Establishment of three-dimensional cultures of human pancreatic duct epithelial cells.
Three-dimensional (3D) cultures of epithelial cells offer singular advantages for studies of morphogenesis or the role of cancer genes in oncogenesis. In this study, as part of establishing a 3D culture system of pancreatic duct epithelial cells, we compared human pancreatic duct epithelial cells (HPDE-E6E7) with pancreatic cancer cell lines. Our results show, that in contrast to cancer cells, HPDE-E6E7 organized into spheroids with what appeared to be apical and basal membranes and a luminal space. Immunostaining experiments indicated that protein kinase Akt was phosphorylated (Ser473) and CTMP, a negative Akt regulator, was expressed in both HPDE-E6E7 and cancer cells. However, a nuclear pool of CTMP was detectable in HPDE-E6E7 cells that showed a dynamic concentrated expression pattern, a feature that further distinguished HPDE-E637 cells from cancer cells. Collectively, these data suggest that 3D cultures of HPDE-E6E7 cells are useful for investigating signaling and morphological abnormalities in pancreatic cancer cells.
Gutierrez-Barrera
Angelica M
AM
Department of Gastrointestinal Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Unit 426, 1515 Holcombe Boulevard, Houston, TX 77030, USA.
Menter
David G
DG
Abbruzzese
James L
JL
Reddy
Shrikanth A G
SA
D021581:Active Transport, Cell Nucleus; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D003094:Collagen / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D004338:Drug Combinations; D004847:Epithelial Cells / Q000166:cytology*; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D010183:Pancreatic Ducts / Q000166:cytology* / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D011509:Proteoglycans / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D014407:Tumor Cells, Cultured / Q000166:cytology*
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D051057:Proto-Oncogene Proteins c-akt