【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。更多还原

【Abstract】 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 models: CNE-1(highly differentiated),CNE-2(poorly differentiated) and the immortalized benign human nasopharyngeal epithelial cell line(NP69) were used in the experiment.Methylation-specific polymerase chain reaction(MS-PCR) assay was performed 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% respectively,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.更多还原