High-level expression of the coxsackievirus and adenovirus receptor messenger RNA in osteosarcoma, Ewing's sarcoma, and benign neurogenic tumors among musculoskeletal tumors.
OBJECTIVE
The sensitivity of human tumor tissues to infection with recombinant adenoviruses correlates with the expression of the coxsackievirus and adenovirus receptor (CAR). CAR has been shown to function as the primary receptor for adenoviruses and to play a critical role in adenovirus entry into host cells. It is important for clinical gene therapy to determine the expression level of CAR in tumor tissues.


METHODS
We analyzed the expression of CAR mRNA in 154 musculoskeletal tumor tissues from 154 patients and 10 normal mesenchymal tissues from 3 patients using reverse transcription-PCR and real-time quantitative PCR. An adenovirus infection assay was performed in two cell lines that were established from CAR-positive osteosarcoma tissue and CAR-negative malignant fibrous histiocytoma tissue.


RESULTS
Ninety-nine of 154 tumors were detected as CAR positive by reverse transcription-PCR. We found that the expression levels of CAR mRNA varied markedly between different tumors as determined by real-time quantitative PCR. CAR mRNA was expressed at high levels in osteosarcoma, Ewing's sarcoma, neurofibroma, and schwannoma; at intermediate levels in exostosis, giant cell tumor, liposarcoma, synovial sarcoma, malignant peripheral nerve sheath tumor, and hemangioma; and at low levels in alveolar soft part sarcoma and desmoid. Whereas the osteosarcoma cell line that expressed a high level of CAR mRNA, like its parent tumor, had a high efficiency of adenovirus infection, the malignant fibrous histiocytoma cell line with almost undetectable expression of CAR mRNA, like its parent tumor, had a low efficiency of infection.


CONCLUSIONS
Our data showed the great variations in CAR mRNA expression among human musculoskeletal tumors and mesenchymal tissues and implicated the potential usefulness of adenoviral vectors in gene therapy for osteosarcoma, Ewing's sarcoma, neurofibroma, and schwannoma. Efficient transduction with adenovirus for gene therapy could be realized in appropriate, sensitive tumor types.
Gu
Wenguang
W
Divisions of Orthopedic Surgery and Cell Biology and Molecular Pharmacology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Ogose
Akira
A
Kawashima
Hiroyuki
H
Ito
Masayuki
M
Ito
Tomoyuki
T
Matsuba
Atsushi
A
Kitahara
Hiroshi
H
Hotta
Tetsuo
T
Tokunaga
Kunihiko
K
Hatano
Hiroshi
H
Morita
Tetsuro
T
Urakawa
Sayuri
S
Yoshizawa
Tatsuya
T
Kawashima
Hiroyuki
H
Kuwano
Ryozo
R
Endo
Naoto
N
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D006367:HeLa Cells; D006801:Humans; D008648:Mesoderm / Q000378:metabolism; D017599:Neuroectodermal Tumors / Q000378:metabolism*; D012516:Osteosarcoma / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D011991:Receptors, Virus / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012512:Sarcoma, Ewing / Q000378:metabolism*; D013997:Time Factors
C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D012333:RNA, Messenger; D011991:Receptors, Virus