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Establishment and characterization of a new human pancreatic adenocarcinoma cell line with high metastatic potential to the lung.

Kalinina T, Güngör C, Thieltges S, Möller-Krull M, Penas EM, Wicklein D, Streichert T, Schumacher U, Kalinin V, Simon R, Otto B, Dierlamm J, Schwarzenbach H, Effenberger KE, Bockhorn M, Izbicki JR, Yekebas EF - BMC Cancer (2010)

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General, Visceral and Thoracic Surgery, University Hospital Hamburg, Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany. tkalinin@uke.de

ABSTRACT

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.

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Related in: MedlinePlus

Whole-genome DNA profile of PaCa 5061. The DNA profile performed by Affymetrix GeneChip hybridization shows a considerable number of large chromosomal alterations. Detailed candidate regions of gene amplification and affected genes are listed in Table 1.
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Figure 4: Whole-genome DNA profile of PaCa 5061. The DNA profile performed by Affymetrix GeneChip hybridization shows a considerable number of large chromosomal alterations. Detailed candidate regions of gene amplification and affected genes are listed in Table 1.

Mentions: The Affymetrix 6.0 SNP array was used to identify the genes amplified in PaCa 5061 cells and primary tumor DNA. The DNA profile of PaCa 5061 cells and the primary tumor of the same patient are shown in Figure 4. Although there are much stronger DNA copy number variations (CNV) in the cell line as compared to the primary tumor sample, the general patterns of CNV were highly similar. The DNA profile showed a considerable number of large chromosomal alterations, including deletions involving at least parts of chromosomes 1p, 2p, 3p, 7q, 8p, 9p, 10, 12q, 17p, 18, and 21. In addition, gains were found at chromosomes 8q, 12p, 14, 16, 17q, 4 and 22. Chromosomal areas of high level gene amplification were found at 2p22, cen6, 7q21, 10p12, 10q11, 12q13, 12q14, and 18q11. The exact amplicon positions and affected genes are summarized in Table 1.


Establishment and characterization of a new human pancreatic adenocarcinoma cell line with high metastatic potential to the lung.

Kalinina T, Güngör C, Thieltges S, Möller-Krull M, Penas EM, Wicklein D, Streichert T, Schumacher U, Kalinin V, Simon R, Otto B, Dierlamm J, Schwarzenbach H, Effenberger KE, Bockhorn M, Izbicki JR, Yekebas EF - BMC Cancer (2010)

Whole-genome DNA profile of PaCa 5061. The DNA profile performed by Affymetrix GeneChip hybridization shows a considerable number of large chromosomal alterations. Detailed candidate regions of gene amplification and affected genes are listed in Table 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927995&req=5

Figure 4: Whole-genome DNA profile of PaCa 5061. The DNA profile performed by Affymetrix GeneChip hybridization shows a considerable number of large chromosomal alterations. Detailed candidate regions of gene amplification and affected genes are listed in Table 1.
Mentions: The Affymetrix 6.0 SNP array was used to identify the genes amplified in PaCa 5061 cells and primary tumor DNA. The DNA profile of PaCa 5061 cells and the primary tumor of the same patient are shown in Figure 4. Although there are much stronger DNA copy number variations (CNV) in the cell line as compared to the primary tumor sample, the general patterns of CNV were highly similar. The DNA profile showed a considerable number of large chromosomal alterations, including deletions involving at least parts of chromosomes 1p, 2p, 3p, 7q, 8p, 9p, 10, 12q, 17p, 18, and 21. In addition, gains were found at chromosomes 8q, 12p, 14, 16, 17q, 4 and 22. Chromosomal areas of high level gene amplification were found at 2p22, cen6, 7q21, 10p12, 10q11, 12q13, 12q14, and 18q11. The exact amplicon positions and affected genes are summarized in Table 1.

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of General, Visceral and Thoracic Surgery, University Hospital Hamburg, Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany. tkalinin@uke.de

ABSTRACT

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.

Show MeSH
Related in: MedlinePlus