鼻咽癌细胞株Syk基因启动子甲基化的研究  

Methylation of Syk gene in promoter region and Syk gene expression in nasopharyngeal carcinoma cell lines

作　　者：鄢冲[1] 金巧智[1] 李志海[2] 刘池波[2] 陶宝鸿[2] 蔡志毅[2] 

机构地区：[1]温州医学院第一临床医学院耳鼻咽喉-头颈外科,浙江温州325000 [2]浙江省台州市立医院耳鼻咽喉-头颈外科,浙江台州318000

出　　处：《中国卫生检验杂志》2012年第2期233-236,共4页Chinese Journal of Health Laboratory Technology

基　　金：浙江省台州市科技计划项目(08KY23)

摘　　要：目的:探讨鼻咽癌细胞株Syk基因启动子甲基化与其mRNA和蛋白质表达情况之间的关系。方法:培养鼻咽癌细胞株CNE-1(高分化)、CNE-2(低分化)和永生化非癌性人鼻咽黏膜上皮细胞株(NP69),采用MS-PCR和Q-RT-PCR及Western Blot方法检测各细胞株中Syk基因的甲基化和Syk mRNA及Syk蛋白表达。结果:MS-PCR法检出CNE-1和CNE-2细胞Syk启动子甲基化率分别为36%±3.6%和62%±4.5%,而NP69未检测到甲基化;Q-RT-PCR法检出Syk mRNA在CNE-1和CNE-2的表达水平均低于NP69细胞,分别为42%±3.5%和28%±2%;WesternBlot法检出Syk蛋白在CNE-1和CNE-2的表达水平均低于NP69细胞,分别为36%±4.5%和16%±2.5%。均有统计学意义(P<0.01)。结论:在分化程度较低的鼻咽癌细胞中,Syk基因启动子甲基化程度较高,则Syk基因mRNA与蛋白的表达较低。Syk基因mRNA和蛋白的表达与Syk基因启动子甲基化程度呈负相关。Objective：To explore the relationship between the methylation of Syk （spleen tyrosine kinase） gene in promoter region and the expression of Syk gene and mRNA in nasopharyngeal carcinoma （NPC） cell lines. Methods： Three cell line mod- els： CNE - 1 （highly differentiated）, CNE -2（ poorly differentiated） and the immortalized benign human nasopharyngeal epitheli- al cell line （NP69） were used in the experiment. Methylation - specific polymerase chain reaction （MS - PCR） assay was per- formed to determine the methylation of Syk gene in promoter region. Quantitative real - time polymerase chain reaction （ Q - RT - PCR） and Western Blot were used to analyze the Syk gene expression in all cell lines. Results： No methylation of Syk promoter was found in NP69 cell line; whereas the rates of Syk promoter methylation in CNE - 1 and CNE - 2 were 36% ＋ 3.6% and 62% ＋4.5% respectively. The Syk mRNA transcription levels in CNE - 1 and CNE - 2 were 42% ＋ 3.5% and 28% - 2% respec- tively, both significantly lower than that in NP69 （P 〈0.01 ）. Also, in CNE -1 and CNE -2, the Syk protein expression had significantly lower levels （36%- 4.5% and 16% - 2.5%, respectively） than that in NP69 （P 〈 0.01 ）. Conclusion： While the rate of Syk promoter methylation was negatively correlated to the differentiation status of NPC cell line; it appeared that Syk promoter methylation and Syk expression in NPC cell lines also had negative correlation.

关 键 词：鼻咽癌 SYK基因 甲基化 启动子 

分 类 号：R739.63[医药卫生—肿瘤]