Jianning Yin1, Qingyu Fan, Xinbao Hao, Degang Fan. 1. Orthopedics Oncology Institute of Chinese PLA, Tangdu Hospital, Fourth Military University, Xi'an, Shanxi, 710038 P. R. China. yinjning@pub.xaonline.com
Abstract
OBJECTIVE: To identify genes associated with metastasis suppression and to investigate the molecular mechanism of osteosarcoma metastasis. METHODS: The subtracted cDNA library of low metastatic human osteosarcoma cell line SOSP-9607 was constructed using suppression subtractive hybridization. Partial clones were sequenced. The acquired sequence data were aligned against the GenBank nucleotide database using Blastn to search for sequence matches. The interested clone was used to perform Northern blot and reverse transcriptase-PCR (RT-PCR) analysis on mRNA isolated from low metastatic cell line SOSP-9607 and OS-9901, high metastatic cell line SOSP-M and three pulmonic metastatic nodules of nude mice. RESULTS: A cDNA clone from low metastatic cell line SOSP-9607 subtracted cDNA library was identified as telomeric repeat binding factor 2 (TERF2) by sequence analysis and Blastn search. Northern blot and RT-PCR analysis demonstrated that TERF2 expressed highly in low metastatic cell line SOSP-9607 and OS-9901, but not in high metastatic cell line SOSP-M and three pulmonic metastatic nodules. CONCLUSION: TERF2 may be important for suppressing metastasis of osteosarcoma.
OBJECTIVE: To identify genes associated with metastasis suppression and to investigate the molecular mechanism of osteosarcoma metastasis. METHODS: The subtracted cDNA library of low metastatic humanosteosarcoma cell line SOSP-9607 was constructed using suppression subtractive hybridization. Partial clones were sequenced. The acquired sequence data were aligned against the GenBank nucleotide database using Blastn to search for sequence matches. The interested clone was used to perform Northern blot and reverse transcriptase-PCR (RT-PCR) analysis on mRNA isolated from low metastatic cell line SOSP-9607 and OS-9901, high metastatic cell line SOSP-M and three pulmonic metastatic nodules of nude mice. RESULTS: A cDNA clone from low metastatic cell line SOSP-9607 subtracted cDNA library was identified as telomeric repeat binding factor 2 (TERF2) by sequence analysis and Blastn search. Northern blot and RT-PCR analysis demonstrated that TERF2 expressed highly in low metastatic cell line SOSP-9607 and OS-9901, but not in high metastatic cell line SOSP-M and three pulmonic metastatic nodules. CONCLUSION:TERF2 may be important for suppressing metastasis of osteosarcoma.