肺癌细胞中NF-κB2基因表达及启动子区域甲基化状态分析  

Analysis of methylation status in promoter region of NF-κB2 and its expression in lung cancer cell lines

作　　者：蒋文斌[1] 李时飞[1] 王丰[1] 许雪青[2] 杨绍俊[1] 贾双荣[1] 李发科[1] 商亚[1] 陈鸣[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所检验科,重庆400042 [2]第三军医大学大坪医院第七研究室,重庆400042

出　　处：《免疫学杂志》2014年第5期432-435,共4页Immunological Journal

基　　金：国家自然科学基金(21107140);重庆市自然科学基金(CSTC2011BB5041)

摘　　要：目的研究肺癌细胞和人正常支气管细胞(human bronchial epithelial cell,HBE)中NF-κB2的表达情况及其启动子区CpG岛的甲基化状态。方法采用实时荧光定量PCR(Real-time PCR)方法检测肺癌细胞和正常支气管细胞NF-κB2基因mRNA的表达情况;采用Western blot技术检测NF-κB2前体蛋白p100的表达情况;采用重亚硫酸盐测序(Bisulfite Sequencing PCR,BSP)技术检测NF-κB2启动子区CpG岛的甲基化状态。结果肺腺癌细胞株A549、肺鳞癌细胞株SK-MES-1和小细胞肺癌细胞株NCI-H446的NF-κB2 mRNA的表达分别是HBE细胞株的6.42±0.91倍(P<0.05,t=5.828)、2.78±0.52倍(P<0.05,t=3.219)、2.79±0.33倍(P<0.05,t=4.611);3种肺癌细胞中p100蛋白较HBE细胞均上调表达;4种细胞株中所扩增片段的CpG位点均呈非甲基化状态。结论肺癌细胞及HBE细胞中NF-κB2启动子区域的CpG位点处于非甲基化状态,说明NF-κB2基因表达情况与甲基化状态无直接关系,其启动子区的甲基化修饰未直接参与该基因的表达调控,可能存在其他相关的调控机制,有待进一步的探索。To investigate the association between expression and methylation status of NF-kB2, we assessed the expression of NF-kB2 and also analyzed its methylation status in three kinds of lung cancer cell lines （A549, SK-MES-1, NCI-H446 ） and HBE cell line. Quantitative real-time PCR and Western blot were used to detect the expression of NF-kB2. In the same time, Bisulfite-sequencing PCR （BSP） was performed to detect the methylation status in the promoter region of NF-kB2. As a consequence, we found that both mRNA and protein expression of NF-kB2 in 3 lung cancer cell lines increased compared with HBE cell line. However, no methylation site of NF- kB2 was found in all the cell lines. Taken these findings together, we may conclude that overexpression of NF-kB2 in lung cancer cell lines is not directly regulated by its promoter methylation. Thus other related regulation mechanism of NF-kB2 overexpression in lung cancer cells should be taken into account further.

关 键 词：NF-kB2 肺肿瘤 甲基化 Bisulfite-sequencing PCR(BSP) 

分 类 号：R734.2[医药卫生—肿瘤] R394.2[医药卫生—临床医学]