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KMID : 0361620090440030285
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Journal of the Korean Orthopaedic Association
2009 Volume.44 No. 3 p.285 ~ p.293
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Aberrant CpG Island Hypermethylation of the RUNX3 Gene in Osteosarcoma
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Oh Joo-Han
Kim Han-Soo
Han Il-Kyu
Kang Gyeong-Hoon
Kim Jun-Hyuk
Cho Hwan-Seong
Kang Seung-Cheol
Lee Mi-Ra
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Abstract
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Purpose: Transcriptional silencing of tumor suppressor genes by aberrant methylation of CpG islands plays a crucial role in the development of human cancers. We comprehensively examined the methylation status of several tumor suppressor genes in osteosarcoma with a special focus on the RUNX3 gene.
Materials and Methods: Methylation-specific polymerase chain reaction (MSP) was performed for osteosarcoma tissues and their cell lines. MSP and RT-PCR for the RUNX3 gene were performed in the tumor-derived cell lines and the immortalized cell lines. The demethylating agent 5-aza-2’ deoxycytidine was used in the SaOS-2 cell line to reverse the methylation status.
Results: Hypermethylation of the RUNX3 gene was observed in 60% (24 of 40) of the osteosarcoma tissues, whereas other tumor suppressor genes showed very low methylation. Thirteen of 30 (43%) tumor-derived cell lines, and U-2OS and SaOS-2 showed hypermethylation of the RUNX3 gene on MSPCR. However, RUNX3 was expressed in the SaOS-2 cell line, as determined by RT-PCR, and the expression was augmented by treatment with 5-aza-2’ deoxycytidine.
Conclusion: Our study suggests that aberrant methylation is an important mechanism of RUNX3 down-regulation in osteosarcoma. This data may have potential significance in developing a potential therapeutic target for osteosarcoma.
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KEYWORD
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RUNX3, Osteosarcoma, Methylation, CpG island
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