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Characterization of human multicentric osteosarcoma using newly established cells derived from multicentric osteosarcoma

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Abstract

Purpose

Human multicentric osteosarcoma (HMOS) is a rare, aggressive variant of osteosarcoma, and its etiology is not clear. We used newly established HMOS cells, which were derived from primary (HMOS-A) and secondary (HMOS-P) lesions, respectively, to perform a basic study analyzing the cellular biology and gene expression of HMOS.

Methods

We performed a cell growth assay, an invasion assay, DNA microarray analysis, quantitative real-time RT–PCR (Qrt-PCR), and a telomerase assay and compared the results between HMOS-A, HMOS-P, and human osteosarcoma (HOS) cell lines (MNNG-HOS and Saos-2).

Results

The cell biological analysis revealed that HMOS-A and HMOS-P had similar characteristics to Saos-2, and the invasion assay showed that they had similar characteristics to MNNG-HOS. The DNA microarray study showed that the gene expression profiles of HMOS-A and HMOS-P were similar to that of MNNG-HOS, but the overexpression of MMP-2, MMP-9, and MT1-MMP was observed in HMOS-A and HMOS-P, which was correlated with the invasiveness of the extracellular matrix, and collagen type-4 (COL-4) and VEGF were also detected. HMOS-A and HMOS-P showed low telomerase activity similar to Saos-2, which are known to be telomerase negative, but a similar telomere length and telomerase protein to MNNG-HOS.

Conclusions

HMOS-A and HMOS-P demonstrated strong invasive ability, and their gene expression profiles correlated with the invasiveness of the extracellular matrix. Their telomerase activity was low, but they did not shown the typical features of alternative lengthening of telomeres (ALT). HMOS-A and HMOS-P are useful models for further study of various biological aspects and therapeutic manipulation of HMOS.

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Abbreviations

ALT:

Alternative lengthening of telomeres

COL-4:

Collagen type-4

GAPDH:

Glyceraldehyde-3 phosphate dehydrogenase

HMOS:

Human multicentric osteosarcoma

HMOS-A:

HMOS Anterior

HMOS-P:

HMOS Posterior

HOS:

Human osteosarcoma

hTERT:

Human telomerase reverse transcriptase

MMP:

Matrix metalloproteinase

MT1-MMP:

Membrane type-1 matrix metalloproteinase

PVDF:

Polyvinylidene difluoride

qRT–PCR:

Quantitative real-time reverse transcriptase polymerase chain reaction

SDS–PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

VEGF:

Vascular endothelial growth factor

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Authors and Affiliations

  1. Department of Orthopaedics Surgery, School of Medicine, Fujita Health University, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake-city, Aichi, 470-1192, Japan

    Y. Yamamoto, K. Tajima, A. Ohno, Y. Washimi, D. Ishimura, O. Washimi & H. Yamada

  2. Laboratory of Molecular Biology and Histochemistry, Joint Research Laboratory, Fujita Health University, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake-city, Aichi, 470-1192, Japan

    N. Yamamoto

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  1. Y. Yamamoto
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  2. N. Yamamoto
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  3. K. Tajima
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  4. A. Ohno
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  5. Y. Washimi
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  8. H. Yamada
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Correspondence to N. Yamamoto.

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Yamamoto, Y., Yamamoto, N., Tajima, K. et al. Characterization of human multicentric osteosarcoma using newly established cells derived from multicentric osteosarcoma. J Cancer Res Clin Oncol 137, 423–433 (2011). https://doi.org/10.1007/s00432-010-0885-9

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  • DOI: https://doi.org/10.1007/s00432-010-0885-9

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