叉头框C2在鼻咽癌侵袭转移中的作用  

Effect of fork head box C2 expression on invasion and metastasis of human nasopharyngeal carcinoma cells

作　　者：李燕姝[1] 张孝文[1] 卢颖深[1] 宋丽娟[1] 赵小龙[1] 黄佳[1] 

机构地区：[1]广州医科大学附属第一医院,510120

出　　处：《中华生物医学工程杂志》2015年第2期114-118,共5页Chinese Journal of Biomedical Engineering

摘　　要：目的：检测叉头框C2（FOXC2）在不同鼻咽癌细胞株中的表达情况，并探讨FOXC2能否通过调控某些上皮标志物或间质标志物的表达诱导上皮间质转分化（EMT），进而参与鼻咽癌细胞侵袭转移。方法以体外培养的人鼻咽癌细胞株（5-8F、CNE2）和永生化鼻咽上皮细胞NP69为研究对象，分别应用Western免疫印迹、实时荧光定量PCR（RT-QPCR）检测3种细胞FOXC2的表达情况。选择低侵袭潜能的鼻咽癌细胞株CNE2，应用瞬时转染FOXC2的表达质粒过表达FOXC2基因，镜下观察转染前后CNE2细胞形态学改变；Transwell迁移与侵袭实验比较转染前后CNE2细胞侵袭转移能力；并采用Western免疫印迹检测EMT相关标志物E-钙黏蛋白、波形蛋白、N-钙黏蛋白的表达情况。结果 FOXC2 mRNA及蛋白在鼻咽癌细胞株5-8F、CNE2及永生化鼻咽上皮细胞NP69中均有表达，且发现其表达量在NP69细胞中最低，在CNE2细胞中较高，而在5-8F细胞中最高（mRNA：0.07±0.05比0.55±0.05比0.81±0.06，蛋白：0.17±0.05比0.56±0.03比0.84±0.06，均P〈0.05）。鼻咽癌细胞株中， FOXC2 mRNA和蛋白在5-8F细胞中的表达高于在CNE2细胞中的表达（mRNA：0.81±0.06比0.55±0.05，蛋白：0.84±0.06比0.56±0.05，均P〈0.05）。FOXC2过表达可诱导CNE2细胞形态由鹅卵石形变为纺锤形样梭形。FOXC2过表达组CNE2细胞运动迁移和侵袭转移能力较空载组增强（迁移细胞数：310±10比80±9，侵袭细胞数：170±10比112±8，均P〈0.05）。与空载组比较，间质标志物波形蛋白和N-钙黏蛋白表达上调（均P〈0.05），上皮标志物E-钙黏蛋白无明显变化。结论 FOXC2可能通过调控间质标志物波形蛋白和N-钙黏蛋白的表达，诱导鼻咽癌发生EMT，从而促进鼻咽癌侵袭转移。Objective To determine the effects of fork head box C2 （FOXC2） expression in different nasopharyngeal carcinoma cell lines,and to investigate whether FOXC2 can induce epithelial-mesenchymal transition（EMT）by regulating epithelial marker or mesenchymal marker expression,so as to involve in the invasion and metastasis of human nasopharyngeal carcinoma cells. Methods The in vitro cultured human nasopharyngeal carcinoma cell lines（5-8F and CNE2）and immortalized nasopharyngeal epithelial cell line NP69 were included as the subjects in the study. The expressions of FOXC2 in the three cell lines were determined by Western blotting and real-time fluorescence quantitative PCR （RT-QPCR）, respectively. The FOXC2 overexpression plasmids were used to transiently transfect the human nasopharyngeal carcinoma cell lines CNE2 of low invasive potential. The changes of CNE2 cell morphology were observed by inverted microscope before and after the transfection. Transwell migration and invasion assays were used to compare the CNE2 cell invasion and metastasis before and after the transfection, respectively. Western blotting was used to examine the expression of the EMT-related markers E-cadherin , vimentin and N-cadherin. Results The expression of FOXC2 mRNA and protein were found in nasopharyngeal carcinoma cell lines 5-8F and CNE2,and immortalized nasopharyngeal epithelial cell line无nbsp;NP69,with the lowest,high and highest level in NP69,CNE2 and 5-8F cells,respectively（mRNA：0.07± 0.05 vs 0.55 ± 0.05 vs 0.81 ± 0.06,protein：0.17 ± 0.05 vs 0.56 ± 0.03 vs 0.84 ± 0.06,all P〈0.05）. In the nasopharyngeal carcinoma cell lines,the expression of FOXC2 mRNA and protein in 5-8F cells was higher than that in CNE2 cells（mRNA：0.81 ± 0.06 vs 0.55 ± 0.05,protein：0.84 ± 0.06 vs 0.56 ± 0.05,all P〈0.05）. The overexpression of FOXC2 could induce the CNE2 cell morphology from cobblestone-like shape to spindle shape. The cell migration and invasion of CNE2 cells in the FOXC2 overexpression group were enha

关 键 词：叉头框C2 鼻咽肿瘤 侵袭 肿瘤转移 

分 类 号：R73-36[医药卫生—肿瘤]