Fu Guobin, Liu Yan, Yuan Jinsong, Zheng Hongwei, Shi Taiping, Lei Wendong, Xiao Ting, Gao Yanning, Cheng Shujun
Department of Etiology and Carcinogenesis, Cancer Institute (Hosptial), Peking Union Medical College & Chinese Academy of Medical Science, Beijing 100021, China.
Zhongguo Fei Ai Za Zhi. 2009 Apr 20;12(4):273-6. doi: 10.3779/j.issn.1009-3419.2009.04.03.
RAP2B is one of the 50 novel candidate genes cloned from the differential expression cDNA libraries constructed in lung cancer cells. Though RAP2B contains conserved domain and belongs to Ras superfamily, the function of RAP2B in carcinogenesis is still poorly understood. The aim of this study is to explore the roles of RAP2B gene in carcinogenesis.
RT-PCR was applied to examine transcriptional status of RAP2B in the tumor and corresponding adjacent tissues collected from 27 patients with lung squamous cell carcinoma. RAP2B expression plasmid was constructed and transfected into Rat1 cells to evaluate the in vitro transformation ability through colony formation assay. Reporter gene assay was performed to reveal the relationship between RAP2B gene and NF-kappaB pathway.
About 67% (18/27) of tumor tissues show higher mRNA expression than that in the corresponding adjacent normal tissues. Typical transforming focus formation was observed in Rat1 cells which were transfected with RAP2B gene. The reporter gene assay data showed that RAP2B activated NF-kappaB pathway more than 3 folds compared with the mock vector.
RAP2B may be a novel candidate oncogene that plays important roles in carcinogenesis through activation of NF-kappaB pathway.
RAP2B是从肺癌细胞构建的差异表达cDNA文库中克隆出的50个新候选基因之一。尽管RAP2B含有保守结构域且属于Ras超家族,但RAP2B在致癌过程中的功能仍知之甚少。本研究的目的是探讨RAP2B基因在致癌过程中的作用。
应用逆转录聚合酶链反应(RT-PCR)检测27例肺鳞状细胞癌患者肿瘤组织及相应癌旁组织中RAP2B的转录状态。构建RAP2B表达质粒并转染至Rat1细胞,通过集落形成试验评估其体外转化能力。进行报告基因检测以揭示RAP2B基因与核因子κB(NF-κB)信号通路之间的关系。
约67%(18/27)的肿瘤组织mRNA表达高于相应的癌旁正常组织。在转染RAP2B基因的Rat1细胞中观察到典型的转化灶形成。报告基因检测数据显示,与空载体相比,RAP2B激活NF-κB信号通路的能力提高了3倍多。
RAP2B可能是一种新的候选癌基因,通过激活NF-κB信号通路在致癌过程中发挥重要作用。