拮抗凋亡转录因子基因在高转移人成骨肉瘤细胞系中的高表达  

Apoptosis Antagonizing Transcript ion Factor is a Gene Highly Expressed in Highly Metastatic Human Osteosar coma Cell Lines

作　　者：殷剑宁[1] 范清宇[1] 郝新宝[1] 杨宁[1] 

机构地区：[1]第四军医大学唐都医院全军骨肿瘤研究所,陕西西安710038

出　　处：《癌症》2002年第2期127-131,共5页Chinese Journal of Cancer

基　　金：国家自然科学基金资助项目(No.39970835)

摘　　要：背景与目的:成骨肉瘤是青少年中最常见的恶性骨肿瘤,具有易发生肺转移的特点。尽管目前已能够成功地控制原发瘤,但是仍有30%以上的患者在5年内死于肺转移。更好地理解调节转移过程的分子机制,为设计有效的治疗方案提供生物学基础,我们用原位移植的方法建立了人成骨肉瘤细胞系SOSP-9607裸鼠转移模型,用这一模型筛选出了高转移细胞系SOSP-M。本研究旨在通过比较两个细胞系的基因表达水平来克隆与骨肉瘤转移相关的基因,探讨成骨肉瘤转移的分子机制。方法:采用抑制性消减杂交技术建立人成骨肉瘤高转移细胞株SOSP-M的消减文库;用差异筛选技术筛选出阳性克隆;对部分克隆进行测序和同源性分析,用Northern杂交及反转录多聚酶链式反应技术对SOSP-M中表达上调的有意义的克隆在低转移细胞系SOSP-9607、OS-9901、高转移细胞系SOSP-M和裸鼠肺转移结节中的表达情况进行了分析。结果:在高转移细胞株SOSP-M中有2个阳性克隆均与人凋亡对抗转录因子(apoptosisantagonizingtransciptionfactor)同源(99%同源)。Northern杂交及反转录PCR证实这个基因在高转移细胞和裸鼠肺转移结节中高表达,而在低转移人成骨肉瘤细胞系SOSP-9607和OS-9901中表达微弱。结论:对抗凋亡转录因子在促进肿瘤细胞转移中可能起重要作用。Background &Objective:Osteosarcoma is the most common mali gnant bone tumor in adolescents and yound adults.It is characterized by a high propensity for pulmonary meta stasis.In spite of successful contr ol of the primary tumor,death from pulmonary metasta ses occurs in >30%of patients within5years.A better understanding of th e molecular mechanisms that regulate the process of metastasis can provide a biological foundation for the design of more effective therapy.We established a metastatic model in nude mice with the method of orthotopically transplanting huma n osteosarcoma cell line SOSP-9607and selected and isolated SOSP-M with highly metasta tic potential.This study is to clone genes associated with osteosarcoma metastasis and to investigate the molecul ar mechanism of osteosarcoma metastasis by comparing the levels of gene expression between the two cells lines.Methods:Using suppression subtractive hybr idization,the subtracted cDNA library of highly me tastatic human osteosarcoma cell line SOSP-M was constructed.Positive clones were screened by differential screen tec hnique.Partial positive clones were sequenced.The interested upexpre ssed clones in SOSP-M cells were analyzed through Northern blot and RT-PCR for the low me tastatic cell lines SOSP-9607and OS-9901,highly metastatic cell line SOSP-M a nd three pulmonic metastatic nodule s.Results:Two positive cDNA clones from highly metastatic cell line SOSP-M subtrac ted cDNA library were identical(99%homology)to apoptosis antagonizing transcription factor.Northern blot and RT-PCR analysis demonstrated that apoptosis a ntagonizing transcription factor e xpressed highly in high metastatic cell line SOSP-M and three pulmonic metastatic nodules,but not in low metastatic cell line SO SP-9607and OS-9901.Conclusion:Apoptosis antagonizing transcript ion factor may play an important role in p romoting metastasis of osteosarcoma.

关 键 词：成骨肉瘤 抑制性消减杂交技术 肿瘤转移 基因表达 凋亡转录因子基因 

分 类 号：R378.1[医药卫生—病原生物学]