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目的:人假想蛋白FLJ22349基因位于染色体22q13.2上,编码一个229个氨基酸的蛋白。表达谱分析发现多种肿瘤组织中有FLJ22349的表达,预测该基因可能与肿瘤的发生发展有密切关系。本研究旨在用生物信息学方法分析FLJ22349的结构和功能,并初步研究FLJ22349在HEK293及乳腺癌细胞中的作用。方法:运用各种生物信息学数据库初步分析FLJ22349的结构和功能。逆转录聚合酶链反应(RT-PCR)方法分析FLJ22349基因在小鼠正常组织,人癌症细胞株及HEK293中的表达情况。将载有FLJ22349的基因的质粒转染入HEK293,MCF-7,MDA-MB231中,观察该基因的亚细胞定位。MTT检测细胞的生长变化。结果:FLJ22349基因在人类、猩猩、恒河猴、褐家鼠、小鼠和狗中有很高的同源性,但与其它已知的基因无同源性。小鼠正常组织中有FLJ22349表达;乳腺癌细胞株MDA-MB231,肝癌细胞株Bel-7402也有表达;主要定位于细胞核;转染FLJ22349的HEK293,MCF-7细胞株生长曲线显示促进细胞生长(P<0.05)。FLJ22349转染120h后,MTT实验观察到HEK293,MCF-7细胞的增殖率分别为299%,456%;而对照组分别为233%,272%。结论:FLJ22349基因过表达促进HEK293和MCF-7细胞的生长。
Objective: Human hypothetical protein FLJ22349 is located on chromosome 22q13.2 and encodes a 229 amino acid protein. Expression profiling revealed FLJ22349 expression in a variety of tumor tissues, and it is predicted that the gene may be closely related to tumorigenesis. The aim of this study was to analyze the structure and function of FLJ22349 by bioinformatics methods and to investigate the role of FLJ22349 in HEK293 and breast cancer cells. Methods: The structure and function of FLJ22349 were analyzed by using various bioinformatics databases. The expression of FLJ22349 gene in mouse normal tissues, human cancer cell lines and HEK293 cells was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The plasmid containing FLJ22349 gene was transfected into HEK293, MCF-7 and MDA-MB231 and the subcellular localization of the gene was observed. MTT detection of cell growth changes. Results: FLJ22349 gene has high homology in human, orangutan, rhesus, rodent, mouse and dog, but not homology with other known genes. FLJ22349 was expressed in normal tissues of mice; MDA-MB231 breast cancer cells and Bel-7402 hepatocellular carcinoma cells were also expressed; mainly located in the nucleus; growth curve of HEK293 and MCF-7 transfected FLJ22349 cells showed that the growth of P <0.05). After transfected with FLJ22349 for 120h, the proliferation rates of HEK293 and MCF-7 cells were 299% and 456%, respectively, compared with 233% and 272% in the control group respectively. Conclusion: FLJ22349 gene overexpression promotes the growth of HEK293 and MCF-7 cells.