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背景与目的:基因CT120是我们实验室从染色体17p13.3位点获得的一个细胞膜相关新基因,mRNA水平检测结果表明该基因在人正常肺组织中不表达,但在人肺癌细胞系SPC-A-1中高表达。本研究进一步从蛋白水平研究CT120在肺癌及癌旁组织中的差异表达及CT120异位表达(ectopicexpression)与过表达(overexpression)对细胞生长的影响。方法:用生物信息学方法设计15肽,合成并偶联到大分子载体KLH上制成人工免疫原,制备鸡抗CT120多肽抗体;通过Westernblotting方法检测CT120在不同肿瘤细胞系中的表达;通过免疫组织化学技术比较CT120在肺癌及癌旁组织中的差异表达;通过集落形成试验研究CT120异位表达对NIH/3T3细胞生长的影响;通过生长曲线及裸鼠成瘤试验研究CT120过表达对A549细胞生长的影响。结果:获得有效的鸡抗CT120抗体(IgY);CT120蛋白在不同肿瘤细胞系中表达程度不同,并在不同类型的肺癌组织中高表达;CT120基因异位表达显著促进NIH/3T3细胞的生长;同时CT120过表达对A549细胞在体内体外的生长有促进作用。结论:基因CT120与肺癌的发生发展有关,是一个人肺癌相关新基因。
BACKGROUND & OBJECTIVE: Gene CT120 is a new cell membrane-related gene obtained from chromosome 17p13.3 in our laboratory. The mRNA level of CT120 is not expressed in human normal lung tissue. However, in human lung cancer cell line SPC-A -1 high expression. This study further investigated the differential expression of CT120 in lung cancer and its adjacent tissues from the protein level and the effects of ectopicexpression and overexpression of CT120 on cell growth. METHODS: Fifteen peptides were designed by bioinformatics method, synthesized and conjugated to the macromolecule carrier KLH to make artificial immunogens to prepare anti-CT120 antibody. The expression of CT120 in different tumor cell lines was detected by Western blotting. The effect of CT120 ectopic expression on the growth of NIH / 3T3 cells was investigated by colony formation assay. The effect of CT120 overexpression on the proliferation of A549 cells The impact of growth. Results: The effective anti-CT120 antibody (IgY) was obtained. The expression of CT120 protein was different in different tumor cell lines and was highly expressed in different types of lung cancer tissues. The ectopic expression of CT120 gene significantly promoted the growth of NIH / 3T3 cells. Overexpression of CT120 can promote the growth of A549 cells in vivo and in vitro. Conclusion: The gene CT120 is related to the occurrence and development of lung cancer and is a new human lung cancer related gene.