应用mRNA差异显示技术克隆人成骨肉瘤转移相关基因

Screening of tumor metastasis related genes from human osteosarcoma cells by mRNA differential display method

筛选与成骨肉瘤转移抑制相关的基因，探讨成骨肉瘤转移的分子机制．方法：应用改良的mRNA差异显示技术，比较成骨肉瘤细胞系SOSP－96o7及其高转移亚系SOSP－M的mRNA表达差异．克隆差异片断，进行测序．根据测序结果设计特异性引物，RT－PCR及Northernblot证实其表达特异性．结果：从低转移细胞SOSP－96O7中分离出2条特异性表达的片断MSPZ和MSP3，经RT－PCR和North．ernblot鉴定，这两条片断只在SOSP－96o7中表达．计算机分析表明2条片断均具有编码区特征，与Genbank和EMBL数据库比较结果显示，MSPZ与已知基因无明显的同源性；MSP3与新克隆的转录因子ZRP有89％的同源性．结论：MSPZ和MSP3可能与成骨肉瘤的转移抑制相关，对它们的进一步研究可能为新的转移抑制基因的发现奠定基础．

AIM: To explore the mechanism of osteosarcomainvasiveness and metastasis. METHODS: Using improvedtechniques of mRNA differential display, the differences ofgene expression between human osteosarcoma cell line SOSP-96o7 and its higher metastatic subpopulation cells SOSP-Mwere studied. RESULTS: Two differentially expressed se-quence tags, MSP2 and MSP3, were cloned from SOSP-96o7and analyzed. RT-PCR and northern blot verified their spe-cific expression in sosp-96o7. MSP2 was novel while MSP3shared 89% sequence homology with ZRP, which was identi-fied as a recently cloned transcription factor. These sequenceshave been logged in Genebank and the database accessionnumbers in EMBL are AF142579 and AF14258O, respective-ly. CONCLUSION: The expression of MSP2 and MSP3 areassociated with the suppression of metastatic property of os-teosarcoma.