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Molecular characterization of commonly used cell lines for bone tumor research: A trans‐European EuroBoNet effort
Author(s) -
Ottaviano Laura,
Schaefer KarlLudwig,
Gajewski Melanie,
Huckenbeck Wolfgang,
Baldus Stefan,
Rogel Uwe,
Mackintosh Carlos,
de Alava Enrique,
Myklebost Ola,
Kresse Stine H.,
MezaZepeda Leonardo A.,
Serra Massimo,
CletonJansen AnneMarie,
Hogendoorn Pancras C. W.,
Buerger Horst,
Aigner Thomas,
Gabbert Helmut E.,
Poremba Christopher
Publication year - 2010
Publication title -
genes, chromosomes and cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.754
H-Index - 119
eISSN - 1098-2264
pISSN - 1045-2257
DOI - 10.1002/gcc.20717
Subject(s) - cdkn2a , biology , cell culture , cancer research , missense mutation , microbiology and biotechnology , multiplex ligation dependent probe amplification , gene , genetics , mutation , exon
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. 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. © 2009 Wiley‐Liss, Inc.