肝癌相关基因HCAP1的定位克隆和初步功能研究  

Cloning of HCAP1 and studies on its primary function

作　　者：王俊茹[1] 覃文新[1] 任功一[1] 潘勇[1] 何祥火[1] 胡建[1] 万大方[1] 顾健人[1] 

机构地区：[1]上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032

出　　处：《肿瘤》2002年第4期263-267,共5页Tumor

基　　金：国家"8 63"生物高技术资助项目 (批准号Z19 0 2 0 1 0 1);国家重点基础研究发展规划"973"资金资助项目 (批准号G19980 5 12 0 9)

摘　　要：目的　克隆和筛选位于染色体 17p13.3缺失区内的肝癌相关基因。方法　采用定位克隆的策略 ,通过PAC筛库和RACE(rapidamplificationofcDNAends)方法克隆新基因 ;通过多组织膜片Northern杂交分析其组织表达谱 ,Southern杂交分析其在肝癌组织DNA水平上的变化 ,然后通过克隆集落形成实验与MTS法测定细胞生长速度 ,初步筛选其作为肿瘤相关基因的可能性。结果　克隆到一个基因HCAP1(HCC associatedprotein 1,先前又称HC5 6 ) ,GenBank登录号为AF177341。多组织膜片杂交显示该基因在多种组织中表达 ,其中胎盘、肝脏和肌肉中表达较高。Southern杂交结果显示HCAP1基因在肝癌组织中存在缺失 ,比例为 2 5 .3% ;克隆集落形成实验与生长抑制率测定实验初步表明该基因能抑制肝癌细胞株Hep3B细胞的生长 ,抑制率为 31.8%。结论　HCAP1可能与肝癌的发生发展相关 ,是一个新的肝癌相关基因的侯选者。Objective To clone and to screen for novel genes related to the hepatocellular carcinoma(HCC) in the deletion region of chromosome 17p13.3. Methods The gene was isolated by a combination of cDNA library screening with PAC and RACE; The tissue expression pattern of HCAP 1 was showed by MTN blot. The deletion of HCAP 1 in cases of HCC was studied by using Southern blot. The inhibitory effect on the Hep3B cells was observed by the colony formation and MTS method. Results The full length cDNA of HCAP 1 (HCC associated protein 1) in the deletion region of chromosome 17p13.3 was cloned. HCAP 1 was expressed in many tissues by MTN blot analysis. The Southern blot revealed the deletion of HCAP 1 gene in 25.3% (17/67) cases of HCC. The colony formation and MTS method showed HCAP 1 protein could reduce the numbers of colony formation and suppress the growth of Hep3B cells. Conclusion HCAP 1 may be related to the hepatocellular carcinoma.

关 键 词：肝癌 HCAP1基因 基因克隆 基因缺失 

分 类 号：R735.7[医药卫生—肿瘤]