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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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mFish analysis of PaCa 5061 cells. mFish analysis of PaCa 5061 cells. Representative karyotype of PaCa 5061 cells. Multiple numerical and unbalanced structural aberrations were observed in the majority of cells. Note. Whole-chromosome losses of 18, 21 and Y, as well as, gains of chromosomes 11 and 20 are frequently found in pancreatic cancer.
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Figure 3: mFish analysis of PaCa 5061 cells. mFish analysis of PaCa 5061 cells. Representative karyotype of PaCa 5061 cells. Multiple numerical and unbalanced structural aberrations were observed in the majority of cells. Note. Whole-chromosome losses of 18, 21 and Y, as well as, gains of chromosomes 11 and 20 are frequently found in pancreatic cancer.

Mentions: An abnormal hypertriploid complex karyotype was detected in all analyzed metaphases of the PaCa 5061 cell line (Figure 3). The karyotype was as follows: 78 < 3n+ > ,XX,-Y,der(1)t(1;9)(p35;?),+2,del(2)(p21)x2,+3,der(3)t(3;12)(p11;?)x2,+5,+7,del(7)(q22)x2,der(8)t(8;16)(p11;?),+del(9)(p13),del(9)(p13),der(9;10)(q10;q10)x2,10,der(10)t(10;16)(q11;?)x2,+11,+12,del(12)(q15)x2,+14,+15,+17,der(17)t(8;17)(?;p11)x2,-18,+19,+20,-21,+22[cp25]. Almost all chromosomes were affected by numerical or structural changes. Numerical abnormalities were found more frequently than structural rearrangements, and chromosomal gains were more frequent than losses. Six unbalanced translocations involving chromosomes 1, 3, 8, 9, 10, and 17 were seen, however, balanced translocations were absent. The identifiable breakpoints of the unbalanced translocations were located in the chromosomal regions 1p35, 3p11, 8p11, 9q10, 10q10/q11, and 17p11. Whole chromosome gains of chromosomes 5, 11, 14, 15, 20, and 22 as well as partial chromosome gains of 2, 3, 7, 8, 9, 12, 16, and 17 were observed in this cell line. Whole chromosome losses affected chromosomes Y, 10, 18, and 21, and partial losses were detected for chromosomes 2, 3, 7, 8, 9, 12, and 17.


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)

mFish analysis of PaCa 5061 cells. mFish analysis of PaCa 5061 cells. Representative karyotype of PaCa 5061 cells. Multiple numerical and unbalanced structural aberrations were observed in the majority of cells. Note. Whole-chromosome losses of 18, 21 and Y, as well as, gains of chromosomes 11 and 20 are frequently found in pancreatic cancer.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: mFish analysis of PaCa 5061 cells. mFish analysis of PaCa 5061 cells. Representative karyotype of PaCa 5061 cells. Multiple numerical and unbalanced structural aberrations were observed in the majority of cells. Note. Whole-chromosome losses of 18, 21 and Y, as well as, gains of chromosomes 11 and 20 are frequently found in pancreatic cancer.
Mentions: An abnormal hypertriploid complex karyotype was detected in all analyzed metaphases of the PaCa 5061 cell line (Figure 3). The karyotype was as follows: 78 < 3n+ > ,XX,-Y,der(1)t(1;9)(p35;?),+2,del(2)(p21)x2,+3,der(3)t(3;12)(p11;?)x2,+5,+7,del(7)(q22)x2,der(8)t(8;16)(p11;?),+del(9)(p13),del(9)(p13),der(9;10)(q10;q10)x2,10,der(10)t(10;16)(q11;?)x2,+11,+12,del(12)(q15)x2,+14,+15,+17,der(17)t(8;17)(?;p11)x2,-18,+19,+20,-21,+22[cp25]. Almost all chromosomes were affected by numerical or structural changes. Numerical abnormalities were found more frequently than structural rearrangements, and chromosomal gains were more frequent than losses. Six unbalanced translocations involving chromosomes 1, 3, 8, 9, 10, and 17 were seen, however, balanced translocations were absent. The identifiable breakpoints of the unbalanced translocations were located in the chromosomal regions 1p35, 3p11, 8p11, 9q10, 10q10/q11, and 17p11. Whole chromosome gains of chromosomes 5, 11, 14, 15, 20, and 22 as well as partial chromosome gains of 2, 3, 7, 8, 9, 12, 16, and 17 were observed in this cell line. Whole chromosome losses affected chromosomes Y, 10, 18, and 21, and partial losses were detected for chromosomes 2, 3, 7, 8, 9, 12, and 17.

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