Identification of candidate liver tumor suppressor genes from human 11p11.2-p12.
Identification of candidate liver tumor suppressor genes from human 11p11.2-p12.
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从人 11p11.2-p12 中鉴定候选肝脏肿瘤抑制基因。
DOI：
10.1002/gcc.1210
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发表时间：
2002
期刊：
影响因子：
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通讯作者：
Coleman,WilliamB
中科院分区：
文献类型：
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作者：
Ricketts,SharonL;Garcia,NicoleF;Betz,BryanL;Coleman,WilliamB
我们之前已经描述了一个用于鉴定人肝肿瘤抑制基因的功能模型，在该模型中，通过微细胞介导的染色体转移将人染色体11导入大鼠肝上皮肿瘤细胞系，产生在体内表现出致瘤性抑制的微细胞杂交(MCH)细胞系。染色体缺失定位研究发现11p11.2-p12的950kb区域保留在所有受抑制的MCH细胞系中，提示该区域可能含有一个或多个具有肝肿瘤抑制功能的基因。在这项研究中，我们通过对一组受抑制的MCH细胞系中142个表达序列标签(EST)标记的检测，构建了11p11.2-p12肝癌抑制区的全面转录图谱。在所检测的142个EST中，19个定位于11p11.2-p12肝癌抑制区。对抑制的MCH细胞系(n=3)中这19个EST的基因表达进行RT-PCR分析，确定了11个潜在的候选肝肿瘤抑制基因。在另外6个受抑制的MCH细胞系中对候选基因表达的检测将潜在候选基因的数量减少到3个(stSG30184、stSG10014和stSG29748)。对受抑制的MCH细胞系和衍生的肿瘤细胞系的Northern印迹分析表明，stSG30184是最好的候选肝癌抑制基因。3.7kb的stSG30184转录本在所有受抑制的MCH细胞系中都有表达，但随着这些细胞的致瘤性的重新表达，其表达被抑制，这与肿瘤抑制基因的表达一致。随后对该EST的鉴定表明，它是一种在多种组织类型中表达的新转录本。这项研究中确定的基因的进一步特征将有助于更好地了解它们在肿瘤性肝病的分子发病机制中的作用。
We have previously described a functional model for identification of human liver tumor suppressor genes in which human chromosome 11 was introduced into rat liver epithelial tumor cell lines via microcell‐mediated chromosome transfer, producing microcell hybrid (MCH) cell lines that exhibit suppression of tumorigenicity in vivo. Chromosome deletion mapping studies identified a 950‐kb region of 11p11.2‐p12 that was retained in all suppressed MCH cell lines, suggesting that this region may harbor one or more genes with liver tumor suppressor function. In this study, we generated a comprehensive transcription map of the 11p11.2‐p12 liver tumor suppressor region through examination of 142 expressed sequence tag (EST) markers among a group of suppressed MCH cell lines. Of 142 ESTs examined, 19 were localized within the 11p11.2‐p12 liver tumor suppressor region. RT‐PCR analysis of gene expression for these 19 ESTs among an index panel of suppressed MCH cell lines (n = 3) identified 11 potential candidate liver tumor suppressor genes. Examination of candidate gene expression among six additional suppressed MCH cell lines reduced the number of potential candidate genes to three (stSG30184, stSG10014, and stSG29748). Northern blot analysis of suppressed MCH cell lines and derived tumor cell lines suggested stSG30184 as the best candidate liver tumor suppressor gene. The 3.7 kb stSG30184 transcript was expressed by all suppressed MCH cell lines, but expression was extinguished coordinately with reexpression of tumorigenicity by these cells, consistent with a tumor suppressor gene. Subsequent characterization of this EST indicates that it is a novel transcript with expression in a broad range of tissue types. Further characterization of the genes identified in this study will provide a greater understanding of their role in the molecular pathogenesis of neoplastic liver disease.