Abstract
Usage of cancer cell lines has repeatedly generated conflicting results provoked by differences among subclones or contamination with mycoplasm or other immortal mammalian cells. To overcome these limitations, we decided within the EuroBoNeT consortium to characterize a common set of cell lines including osteosarcomas (OS), Ewing sarcomas (ES), and chondrosarcomas (CS). DNA fingerprinting was used to guarantee the identity of all of the cell lines and to distinguish subclones of osteosarcoma cell line HOS. Screening for homozygous loss of 38 tumor suppressor genes by MLPA revealed deletion of CDKN2A as the most common event (15/36), strictly associated with absence of the CDKN2A (p16) protein. Ten cell lines showed missense mutations of the TP53 gene while another set of nine cell lines showed mutations resulting in truncation of the TP53 protein. Cells harboring missense mutations expressed high levels of nuclear TP53, while cell lines with nonsense mutations showed weak/absent staining for TP53. TP53wt cell lines usually expressed the protein in 2-10% of the cells. However, seven TP53wt osteosarcomas were negative for both mRNA and protein expression. Our analyses shed light on the correlation between immunohistochemical and genetic data for CDKN2A and TP53, and confirm the importance of these signaling pathways. The characterization of a substantial number of cell lines represents an important step to supply research groups with proven models for further advanced studies on tumor biology and may help to make results from different laboratories more comparable.
| Original language | English |
|---|---|
| Pages (from-to) | 40-51 |
| Number of pages | 12 |
| Journal | Genes Chromosomes and Cancer |
| Volume | 49 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jan 2010 |
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ASJC Scopus subject areas
- Cancer Research
- Genetics
Cite this
Molecular characterization of commonly used cell lines for bone tumor research : A trans-European EuroBoNet effort. / Ottaviano, Laura; Schaefer, Karl Ludwig; Gajewski, Melanie; Huckenbeck, Wolfgang; Baldus, Stefan; Rogel, Uwe; Mackintosh, Carlos; De Alava, Enrique; Myklebost, Ola; Kresse, Stine H.; Meza-Zepeda, Leonardo A.; Serra, Massimo; Cleton-Jansen, Anne Marie; Hogendoorn, Pancras C W; Buerger, Horst; Aigner, Thomas; Gabbert, Helmut E.; Poremba, Christopher.
In: Genes Chromosomes and Cancer, Vol. 49, No. 1, 01.2010, p. 40-51.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Molecular characterization of commonly used cell lines for bone tumor research
T2 - A trans-European EuroBoNet effort
AU - Ottaviano, Laura
AU - Schaefer, Karl Ludwig
AU - Gajewski, Melanie
AU - Huckenbeck, Wolfgang
AU - Baldus, Stefan
AU - Rogel, Uwe
AU - Mackintosh, Carlos
AU - De Alava, Enrique
AU - Myklebost, Ola
AU - Kresse, Stine H.
AU - Meza-Zepeda, Leonardo A.
AU - Serra, Massimo
AU - Cleton-Jansen, Anne Marie
AU - Hogendoorn, Pancras C W
AU - Buerger, Horst
AU - Aigner, Thomas
AU - Gabbert, Helmut E.
AU - Poremba, Christopher
PY - 2010/1
Y1 - 2010/1
N2 - Usage of cancer cell lines has repeatedly generated conflicting results provoked by differences among subclones or contamination with mycoplasm or other immortal mammalian cells. To overcome these limitations, we decided within the EuroBoNeT consortium to characterize a common set of cell lines including osteosarcomas (OS), Ewing sarcomas (ES), and chondrosarcomas (CS). DNA fingerprinting was used to guarantee the identity of all of the cell lines and to distinguish subclones of osteosarcoma cell line HOS. Screening for homozygous loss of 38 tumor suppressor genes by MLPA revealed deletion of CDKN2A as the most common event (15/36), strictly associated with absence of the CDKN2A (p16) protein. Ten cell lines showed missense mutations of the TP53 gene while another set of nine cell lines showed mutations resulting in truncation of the TP53 protein. Cells harboring missense mutations expressed high levels of nuclear TP53, while cell lines with nonsense mutations showed weak/absent staining for TP53. TP53wt cell lines usually expressed the protein in 2-10% of the cells. However, seven TP53wt osteosarcomas were negative for both mRNA and protein expression. Our analyses shed light on the correlation between immunohistochemical and genetic data for CDKN2A and TP53, and confirm the importance of these signaling pathways. The characterization of a substantial number of cell lines represents an important step to supply research groups with proven models for further advanced studies on tumor biology and may help to make results from different laboratories more comparable.
AB - Usage of cancer cell lines has repeatedly generated conflicting results provoked by differences among subclones or contamination with mycoplasm or other immortal mammalian cells. To overcome these limitations, we decided within the EuroBoNeT consortium to characterize a common set of cell lines including osteosarcomas (OS), Ewing sarcomas (ES), and chondrosarcomas (CS). DNA fingerprinting was used to guarantee the identity of all of the cell lines and to distinguish subclones of osteosarcoma cell line HOS. Screening for homozygous loss of 38 tumor suppressor genes by MLPA revealed deletion of CDKN2A as the most common event (15/36), strictly associated with absence of the CDKN2A (p16) protein. Ten cell lines showed missense mutations of the TP53 gene while another set of nine cell lines showed mutations resulting in truncation of the TP53 protein. Cells harboring missense mutations expressed high levels of nuclear TP53, while cell lines with nonsense mutations showed weak/absent staining for TP53. TP53wt cell lines usually expressed the protein in 2-10% of the cells. However, seven TP53wt osteosarcomas were negative for both mRNA and protein expression. Our analyses shed light on the correlation between immunohistochemical and genetic data for CDKN2A and TP53, and confirm the importance of these signaling pathways. The characterization of a substantial number of cell lines represents an important step to supply research groups with proven models for further advanced studies on tumor biology and may help to make results from different laboratories more comparable.
UR - http://www.scopus.com/inward/record.url?scp=74949090294&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=74949090294&partnerID=8YFLogxK
U2 - 10.1002/gcc.20717
DO - 10.1002/gcc.20717
M3 - Article
C2 - 19787792
AN - SCOPUS:74949090294
VL - 49
SP - 40
EP - 51
JO - Genes Chromosomes and Cancer
JF - Genes Chromosomes and Cancer
SN - 1045-2257
IS - 1
ER -