目的克隆并分析新型基因CCP22,研究其结构及初探生物学特性。方法常规分子克隆、生物信息学分析、Westernblot、RT-PCR。结果利用酵母双杂交技术从人乳腺文库中分离到一种含有22个补体调控蛋白(complement control protein,CCP)序列元件的新基因,命名为CCP22。生物信息学分析发现,该基因定位于人染色体9q31.2-32,共15个外显子,基因编码序列全长4494bp,编码1497个氨基酸。将CCP22基因全长编码序列克隆于真核载体中,该表达载体转染人胚肾细胞293T后获得的表达产物经Westernblot证实CCP22属于分泌蛋白。将CCP22与绿色荧光蛋白(EGFP)标签融合后证实CCP22主要分布在细胞核外。Northernblot证实,内源性CCP22mRNA转录本全长约6kb。RT-PCR检测表明,CCP22在正常乳腺细胞株中高表达,而在多种乳腺肿瘤细胞株中不表达或低表达。结论CCP22在乳腺肿瘤发生发展中可能发挥重要作用。
To explore the biological roles of human Pescadillo and investigate its potential effect on tumorigene- sis, the cDNA of Pescadillo was fused with that of GST. After purification and elution, the purified GST-Pescadillo fusion protein was obtained, and the antibody against the fusion protein was generated. Endogenous Pescadillo protein was observed to be remarkably induced by estrogen. It was mainly distributed in the tissues such as breast, ovary and intestine, all of which contain proliferating cells, and was also detected in many cell lines of human cancer: renal carcinoma, hepatoma, ovarian cancer, colon carcinoma, and breast cancer. The expression level of Pescadillo was increased significantly in breast cancer tissues compared with their paired margin tissues. Taken together, these data suggest that Pescadillo may play important roles in the initiation and development of cancer and may be a po- tential target in cancer diagnosis and therapy.
ZHANG Hao1, LI JiePing1, WANG XiaoHui1, SUN Yan1, YUAN Bin1, YANG ZhiHong1, JIANG YanChao1, ZENG Min1, DING LiHua1, NING Kang1, ZHU JianHua2, LI JieZhi1, HUANG CuiFen1, LIU AiJun3 & YE QiNong1 1 Beijing Institute of Biotechnology, Beijing 100850, China
