Pan W, Zhang Q, Xi Q S, Gan R B, Li T P
Institute of Biochemistry and Cell Biology, Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai, 200031, China.
Biochem Biophys Res Commun. 2001 Sep 7;286(5):1033-8. doi: 10.1006/bbrc.2001.5478.
Human primary hepatocellular carcinoma (HCC)is one of the highly prevalent malignant diseases worldwide, the identification of HCC-associated genes has been a major approach in elucidating the molecular mechanism of tumorigenesis of HCC. In our previous studies, a function-unknown gene, which displayed marked expression difference between the HCC sample and normal liver control has been detected by cDNA microarray. This gene was named after fup1 (function-unknown protein 1), and was cloned according to the data of GenBank. The cDNA of fup1 has an open-reading frame 1233 base pairs in size. Here, the function analysis of FUP1 related to HCC is being reported. The NIH3T3 cells transiently transfected with FLAG-conjugated FUP1 revealed strong nuclear staining in immunofluorescent assay. Furthermore, cell proliferation enhancing activity of fup1 was shown by MTT assay in stable transfectant NIH3T3 cell line with pcDNA3-derived plasmid having fup1 under the regulation of pCMV, while cell proliferation repressing activity of antisense fup1 was observed in BEL7404 stable transfectant cells. Tumorigenicity of the above stable transfectant cells was analyzed in nude mice compared with appropriate controls. The result was in good agreement with MTT assay. Elevated tumorigenicity of fup1 transfected NIH3T3 cell and repressed tumorigenicity of antisense fup1 transfected BEL7404 cell were clearly demonstrated. The results above suggested that fup1 might be a critical gene related to carcinogenesis of HCC. Detailed molecular function of fup1 remains to be elucidated.
人类原发性肝细胞癌（HCC）是全球高度流行的恶性疾病之一，鉴定HCC相关基因一直是阐明HCC肿瘤发生分子机制的主要方法。在我们之前的研究中，通过cDNA微阵列检测到一个功能未知的基因，该基因在HCC样本和正常肝脏对照之间表现出明显的表达差异。这个基因被命名为fup1（功能未知蛋白1），并根据GenBank的数据进行克隆。fup1的cDNA有一个大小为1233个碱基对的开放阅读框。在此，报告了与HCC相关的FUP1的功能分析。用FLAG标记的FUP1瞬时转染的NIH3T3细胞在免疫荧光测定中显示出强烈的核染色。此外，在具有pCMV调控下的fup1的pcDNA3衍生质粒的稳定转染NIH3T3细胞系中，MTT测定显示fup1具有细胞增殖增强活性，而在BEL7404稳定转染细胞中观察到反义fup1的细胞增殖抑制活性。与适当的对照相比，在裸鼠中分析了上述稳定转染细胞的致瘤性。结果与MTT测定结果高度一致。清楚地证明了fup1转染的NIH3T3细胞的致瘤性升高和反义fup1转染的BEL7404细胞的致瘤性降低。上述结果表明，fup1可能是与HCC致癌作用相关的关键基因。fup1的详细分子功能仍有待阐明。
