NID1促进人卵巢癌细胞OVCAR-3侵袭转移及分子机制的研究  

Study of NID1-promoted Migration and Invasion and Its Molecular Mechanism in Ovarian Cancer Cell OVCAR-3

作　　者：朱远远[1,2] 张春冬[1,2] 王义涛[1,2] 王森[1,2] 朱慧芳[1,2] 李轶[1,2] 雷云龙[1,2] 卜友泉[1,2] 张莹[1,2] 

机构地区：[1]重庆医科大学生物化学与分子生物学教研室,重庆400016 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016

出　　处：《中国细胞生物学学报》2015年第2期221-228,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81302263);重庆市科委自然科学基金(批准号:cstc2014jcyj A10026);重庆市教委科学技术研究项目(批准号:KJ130326;KJ1400207)资助的课题~~

摘　　要：Nidogen-1(NID1)是新发现的一个候选卵巢癌诊断标志物,该研究旨在初探其在卵巢癌细胞中的生物学功能。首先构建稳定过表达外源NID1的人卵巢癌细胞系OVCAR-3,然后采用划痕实验和Transwell迁移和侵袭实验检测细胞的迁移侵袭能力,并通过荧光定量PCR和Western blot检测细胞中上皮–间质转化(epithelial-mesenchymal transition,EMT)相关蛋白和ERK/MAPK通路蛋白的表达情况。结果显示,与空载细胞相比,稳定过表达NID1的OVCAR-3细胞其划痕愈合能力、细胞迁移和侵袭能力均明显增强。较之空载细胞的上皮细胞样外形,稳定过表达NID1的OVCAR-3细胞呈间质细胞样外形,其上皮细胞标志分子E-cadherin表达下调,间质细胞标志分子(包括Vimentin和N-cadherin)和EMT相关转录因子Twist-2表达上调。此外,稳定过表达NID1的OVCAR-3细胞中ERK1/2的磷酸化水平升高,经ERK/MAPK通路的抑制剂U0126下调ERK1/2的磷酸化水平后其Ecadherin、Vimentin、N-cadherin和Twist-2表达水平出现逆转。这些结果提示,NID1可能通过激活ERK/MAPK通路促进卵巢癌细胞的EMT过程,进而增强其侵袭转移的能力。Nidogen-1（NID1） is found to be a novel candidate diagnostic biomarker of ovarian cancer in our previous study, but its role in the development of ovarian cancer is unclear. In this study, we firstly established the OVCAR-3 monoclone with stable ectopic expression of NID1, and then assessed its migratory and invasive abilities by wound healing assay, Transwell migration and invasion assays. Subsequently, we utilized quantitative RT-PCR and Western blot to detect proteins relating to epithelial-mesenchymal transition（EMT） and ERK/MAPK signaling pathway in the NID1-overexpressed OVCAR-3 cells. The results showed that NID1-overexpressed OVCAR-3 cells exhibited significantly greater motility and invasiveness comparing with those of the control group; NID1 overexpression also led to the enhanced EMT phenotype, including the fibroblastic morphology, the reduction of the epithelial marker E-cadherin, the enhancement of mesenchymal markers（Vimentin and N-cadherin） and the transcription factor Twist-2. Furthermore, the levels of phosphorylated ERK1/2 were increased in NID1-overexpressed OVCAR-3 cells. After decreasing the ERK1/2 phosphorylation in these cells treated with an MEK inhibitor, U0126, the expressions of EMT relevant marks were regressed. Taken together, our findings reveal that NID1 promotes ovarian cancer metastasis, probably through the activation of the ERK/MAPK signaling pathway to propel EMT.

关 键 词：NID1 卵巢癌 转移 侵袭 EMT 

分 类 号：R737.31[医药卫生—肿瘤]