Further characterization of the first seminoma cell line TCam-2.
Testicular germ cell tumors of adolescents and adults (TGCTs) can be classified into seminomatous and nonseminomatous tumors. Various nonseminomatous cell lines, predominantly embryonal carcinoma, have been established and proven to be valuable for pathobiological and clinical studies. So far, no cell lines have been derived from seminoma which constitutes more than 50% of invasive TGCTs. Such a cell line is essential for experimental investigation of biological characteristics of the cell of origin of TGCTs, i.e., carcinoma in situ of the testis, which shows characteristics of a seminoma cell. Before a cell line can be used as model, it must be verified regarding its origin and characteristics. Therefore, a multidisciplinary approach was undertaken on TCam-2 cells, supposedly the first seminoma cell line. Fluorescence in situ hybridization, array comparative genomic hybridization, and spectral karyotyping demonstrated an aneuploid DNA content, with gain of 12p, characteristic for TGCTs. Genome wide mRNA and microRNA expression profiling supported the seminoma origin, in line with the biallelic expression of imprinted genes IGF2/H19 and associated demethylation of the imprinting control region. Moreover, the presence of specific markers, demonstrated by immunohistochemistry, including (wild type) KIT, stem cell factor, placental alkaline phosphatase, OCT3/4 (also demonstrated by a specific Q-PCR) and NANOG, and the absence of CD30, SSX2-4, and SOX2, confirms that TCam-2 is a seminoma cell line. Although mutations in oncogenes and tumor suppressor genes are rather rare in TGCTs, TCam-2 had a mutated BRAF gene (V600E), which likely explains the fact that these cells could be propagated in vitro. In conclusion, TCam-2 is the first well-characterized seminoma-derived cell line, with an exceptional mutation, rarely found in TGCTs.
Department of Pathology, Daniel den Hoed Cancer Center, Josephine Nefkens Institute, Erasmus MC-University Medical Center Rotterdam, Rotterdam, The Netherlands.
Jeroen
J
de Jong
Hans
H
Stoop
Ad J M
AJ
Gillis
Remko
R
Hersmus
Ruud J H L M
RJ
van Gurp
Gert-Jan M
GJ
van de Geijn
Ellen
E
van Drunen
H Berna
HB
Beverloo
Dominik T
DT
Schneider
Jon K
JK
Sherlock
John
J
Baeten
Sohei
S
Kitazawa
E Joop
EJ
van Zoelen
Kees
K
van Roozendaal
J Wolter
JW
Oosterhuis
Leendert H J
LH
Looijenga
D045744:Cell Line, Tumor; D020869:Gene Expression Profiling; D018392:Genomic Imprinting; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D009363:Neoplasm Proteins / Q000032:analysis; D009693:Nucleic Acid Hybridization; D018239:Seminoma / Q000473:pathology*; D032681:Spectral Karyotyping
D009363:Neoplasm Proteins