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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 investigate the relationship between methylation of Syk gene promoter and its mRNA and protein expression in nasopharyngeal carcinoma cell lines. Methods: Nasopharyngeal carcinoma cell line CNE-1 (well-differentiated), CNE-2 (poorly differentiated) and immortalized nasopharyngeal epithelial cell line (NP69) Western Blot method was used to detect Syk gene methylation and Syk mRNA and Syk protein expression in each cell line. Results: The methylation rates of Syk promoter in CNE-1 and CNE-2 cells were 36% ± 3.6% and 62% ± 4.5%, respectively, but no methylation was detected in NP69 cells by MS-PCR. The expression levels of Syk mRNA in CNE-1 and CNE-2 were lower than those in NP69 cells by PCR (42% ± 3.5% and 28% ± 2%, respectively). Syk protein was detected by Western Blot in CNE-1 and CNE- 2 were all lower than that of NP69 cells (36% ± 4.5% and 16% ± 2.5%, respectively). All were statistically significant (P <0.01). CONCLUSIONS: Syk gene promoter methylation is higher in nasopharyngeal carcinoma cells with lower differentiation, while Syk mRNA and protein expression is lower. Syk mRNA and protein expression was negatively correlated with the degree of methylation of Syk promoter.