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背景与目的RAP2B是我们实验室构建的中国人肺鳞癌差异表达cDNA文库中高表达的基因之一。作为具有保守结构域的Ras超家族成员,RAP2B基因在肿瘤发生发展中的作用仍属未知。本文拟初步探讨RAP2B基因在肺癌中的作用及其机制。方法应用半定量RT-PCR方法检测了27例手术切除的肺鳞癌肿瘤组织和相应的癌旁组织中RAP2B基因的表达;结合生物信息学分析克隆全长ORF区并转染大鼠Rat1细胞,观察转化灶形成情况;采用NF-kappaB信号通路特异的报告基因观察RAP2B基因对NF-kappaB通路的影响。结果RAP2B mRNA在约67%(18/27)的肺鳞癌配对组织中肿瘤较癌旁组织高表达;成功构建了RAP2B的真核表达质粒;平板转化实验显示转染RAP2B基因的细胞出现明显转化灶;通路报告基因分析显示RAP2B能够明显激活NF-kappaB通路。结论RAP2B基因在肺癌患者的肿瘤组织高表达,在体外具有恶性转化Rat1细胞能力,提示其为候选癌基因,并且可能通过激活NF-kappaB信号通路在肺癌发生发展中发挥作用。
BACKGROUND & OBJECTIVE: RAP2B is one of the most highly expressed genes in Chinese squamous cell carcinoma differentially expressed cDNA libraries constructed in our laboratory. As a member of the Ras superfamily with conserved domains, the role of RAP2B in tumorigenesis remains unknown. This article intends to investigate the role of RAP2B gene in lung cancer and its mechanism. Methods The expression of RAP2B gene was detected by semi-quantitative RT-PCR in 27 surgically resected lung squamous cell carcinoma tissues and its corresponding paracancerous tissues. The full-length ORF region was cloned by bioinformatics analysis and transfected into rat Rat1 cells. The formation of foci was observed. The effect of RAP2B gene on the NF-kappaB pathway was observed by using the reporter gene specific NF-kappaB signaling pathway. Results RAP2B mRNA was highly expressed in paired lung squamous cell carcinoma tissues of 67% (18/27) of the squamous cell carcinoma tissues. The eukaryotic expression plasmid of RAP2B gene was constructed successfully. The transformation of RAP2B gene cells showed obvious transformation Pathway reporter analysis showed that RAP2B significantly activated NF-kappaB pathway. Conclusion The RAP2B gene is highly expressed in the tumor tissue of lung cancer patients and has the ability of malignant transformation of Rat1 cells in vitro, suggesting that RAP2B gene is a candidate oncogene and may play a role in the development of lung cancer by activating NF-kappaB signaling pathway.