骨肿瘤转移抑制基因全长cDNA克隆、测序及功能分析

To amplificate the complete cDNAs of MSP2 and MSP3 associated with metastasis suppress genes of osteosarcoma, and to analysis their functions. Method: The total mRNA of osteosarcoma SOSP-9607 was extracted and used to synthesis cDNA. The complete cDNAs of MSP2 and MSP3 were screened, cloned into a eukaryocyte expression vector separately. The osteosarcoma cell SOSP-M was transfected with the reconstructed plasmid, sceened by G418, and then injected nude mouse. The metastasis ability of the transfected cell and untransfected cell was compared. The segments of metastasis associated genes were labeled with digoxin, applied to hybridization in situ, FISH and PRINS studies of different origin tumors with or without metastasis. Result: Two decrease libraries of SOSP-9607 and SOSP-M were constructed by SSH. There was a clone screened homologous to TERF2 (telomeric repeat binding factor 2). It is proved by Northern blot and RT-PCR that the gene only expressed in low metastasis osteosarcoma SOSP-9607, not expressed in high metastasis osteosarcoma SOSP-M, and not in metastasis nods of nude lung. There were 2 clones screened with homogeneity 99% to apoptosis antagonizing transciption factor. It is showed by Northern blot and RT-PCR that the gene expressed highly in metastasis osteosarcoma cell SOSP-M and in metastasis nods of nude lung, expressed lowly in osteosarcoma SOSP-9607 and OS-9901. The whole length gene of MSP2 was amplified by RACE. It is found that the homologous gene of the new gene is hypothetical protein FLJ20062, which coding a function unknown protein, according to the GenBank and dbETS. Conclusion: TERF2 may play an important role in suppression of cell development to metastasis, in maintenance stabilize of tumor cell, and in suppression of osteosarcoma development. The apoptosis antagonizing transciption factor may be important in acceleration of tumor metastasis. The studies of metastasis associated genes will be useful to the basis of gene therapy

在我室已筛选出骨肉瘤转移抑制基因片段的基础上，以Marathon cDNA扩增法筛出全长基因尴赴胩逑导凹扑慊Ｄ夥治銎浞肿恿考肮δ埽还菇ㄕ婧吮泶镌靥澹靖咦乒侨饬鱿赴鄄楹笳叩纳镅形谋洹ISH及PRINS法染色体定位。用原位杂交法评价法评价其表达在不同起源的转移和非转移肿瘤的普遍性意义。