Deng Li, Xiang Xiaocong, Yang Fei, Xiao Dongqin, Liu Kang, Chen Zhu, Zhang Ruolan, Feng Gang
Research Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital, The Second Clinical College of North Sichuan Medical College, Nanchong, Sichuan 637000, China.
Orthopedics, Southwest Medical University, Luzhou, Sichuan 646000, China.
Biochem Biophys Res Commun. 2017 Aug 19;490(2):161-168. doi: 10.1016/j.bbrc.2017.06.016. Epub 2017 Jun 8.
Cancer cell molecular mimicry of stem cells (SC) follows with enhanced proliferative and renewal capacities. In support, numerous mediators of SC self-renewal have been evinced to exhibit oncogenic potential. More and more researches showed that the embryonic stem cell self-renewal genes express in various cancer cells. In this study, we sought to test the tumorigenic functions of NANOG, particularly, in esophageal cancer (EC). Using quantitative RT-PCR and western blotting, we confirmed that EC cells highly express NANOG mRNA and protein. We then constructed a shRNA-mediated plasmid to knockdown of NANOG mRNA. We observed that NANOG deficiency in Eca109 cells decreased clone formation, cell proliferation, and showed G1 arrest. To further investigate the functions and mechanisms of NANOG in Eca109 cells, we detected the changes of multiple signaling molecules when NANOG deficiency. We foud that NANOG deficiency affected multiple genes, particularly, supressed drug-resistance via down-regulated ABCG2 in Eca109 cells, and caused G1 arrest by down-regulated cyclin D1 (CCND1) expression. The present loss-of-function work, establish the integral role for NANOG in Eca109 cell proliferation, drug resistance, and shed light on its mechanisms of action.
癌细胞对干细胞的分子模拟伴随着增殖和更新能力的增强。作为证据，众多干细胞自我更新的介质已被证明具有致癌潜力。越来越多的研究表明，胚胎干细胞自我更新基因在各种癌细胞中表达。在本研究中，我们试图测试NANOG的致瘤功能，特别是在食管癌（EC）中的功能。通过定量逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法，我们证实EC细胞高表达NANOG mRNA和蛋白质。然后我们构建了一个短发夹RNA（shRNA）介导的质粒以敲低NANOG mRNA。我们观察到Eca109细胞中NANOG的缺失减少了克隆形成、细胞增殖，并导致G1期阻滞。为了进一步研究NANOG在Eca109细胞中的功能和机制，我们检测了NANOG缺失时多种信号分子的变化。我们发现NANOG的缺失影响多个基因，特别是在Eca109细胞中通过下调ABCG2抑制耐药性，并通过下调细胞周期蛋白D1（CCND1）的表达导致G1期阻滞。目前的功能丧失研究确定了NANOG在Eca109细胞增殖、耐药性中的整体作用，并揭示了其作用机制。
