Research article
Establishment and characterization of a new human pancreatic adenocarcinoma cell line with high metastatic potential to the lung
-
* Corresponding author: Tatyana Kalinina tkalinin@uke.de
- † Equal contributors
1 Department of General, Visceral and Thoracic Surgery, University Hospital Hamburg, Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
2 Department of Clinical Chemistry, University Hospital Hamburg, Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
3 Hubertus Wald Tumorzentrum, University Cancer Center Hamburg, University Medical Center, University Hospital Hamburg, Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
4 Institute of Anatomy and Experimental Morphology, University Hospital Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
5 Institute of Pathology, University Hospital Hamburg, Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
6 Institute of Tumor Biology, University Hospital Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany
BMC Cancer 2010, 10:295 doi:10.1186/1471-2407-10-295
Published: 16 June 2010Abstract
Background
Pancreatic cancer is still associated with devastating prognosis. Real progress in treatment options has still not been achieved. Therefore new models are urgently needed to investigate this deadly disease. As a part of this process we have established and characterized a new human pancreatic cancer cell line.
Methods
The newly established pancreatic cancer cell line PaCa 5061 was characterized for its morphology, growth rate, chromosomal analysis and mutational analysis of the K-ras, EGFR and p53 genes. Gene-amplification and RNA expression profiles were obtained using an Affymetrix microarray, and overexpression was validated by IHC analysis. Tumorigenicity and spontaneous metastasis formation of PaCa 5061 cells were analyzed in pfp-/-/rag2-/- mice. Sensitivity towards chemotherapy was analysed by MTT assay.
Results
PaCa 5061 cells grew as an adhering monolayer with a doubling time ranging from 30 to 48 hours. M-FISH analyses showed a hypertriploid complex karyotype with multiple numerical and unbalanced structural aberrations. Numerous genes were overexpressed, some of which have previously been implicated in pancreatic adenocarcinoma (GATA6, IGFBP3, IGFBP6), while others were detected for the first time (MEMO1, RIOK3). Specifically highly overexpressed genes (fold change > 10) were identified as EGFR, MUC4, CEACAM1, CEACAM5 and CEACAM6. Subcutaneous transplantation of PaCa 5061 into pfp-/-/rag2-/- mice resulted in formation of primary tumors and spontaneous lung metastasis.
Conclusion
The established PaCa 5061 cell line and its injection into pfp-/-/rag2-/- mice can be used as a new model for studying various aspects of the biology of human pancreatic cancer and potential treatment approaches for the disease.