Molecular characterization of commonly used cell lines for bone tumor research: a trans-European EuroBoNet effort.
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. TP53(wt) cell lines usually expressed the protein in 2-10% of the cells. However, seven TP53(wt) 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.
Ottaviano
Laura
L
Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf, Germany.
Schaefer
Karl-Ludwig
KL
Gajewski
Melanie
M
Huckenbeck
Wolfgang
W
Baldus
Stefan
S
Rogel
Uwe
U
Mackintosh
Carlos
C
de Alava
Enrique
E
Myklebost
Ola
O
Kresse
Stine H
SH
Meza-Zepeda
Leonardo A
LA
Serra
Massimo
M
Cleton-Jansen
Anne-Marie
AM
Hogendoorn
Pancras C W
PC
Buerger
Horst
H
Aigner
Thomas
T
Gabbert
Helmut E
HE
Poremba
Christopher
C
D000818:Animals; D035843:Biomedical Research; D001859:Bone Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D003299:Cooperative Behavior; D019941:Cyclin-Dependent Kinase Inhibitor p16; D005060:Europe; D006801:Humans; D016159:Tumor Suppressor Protein p53
D019941:Cyclin-Dependent Kinase Inhibitor p16; D016159:Tumor Suppressor Protein p53