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利用5-杂氮-2′-脱氧胞苷(5-aza-2′-deoxycytidine,5-aza-CdR)处理体外培养的鼻咽癌细胞株CNE-1、CNE-2及永生化非癌性人鼻咽上皮细胞株NP-69,采用BS-PCR、Q-RT-PCR及Westernblot方法分别检测经5μmol/L的5-aza-CdR处理前后,各细胞株中Syk基因启动子甲基化状况及SykmRNA和蛋白质表达情况。探讨去甲基化药物5-杂氮-2′-脱氧胞苷(5-aza-CdR)对鼻咽癌细胞株中脾酪氨酸激酶(spleen tyrosine kinase,Syk)启动子甲基化水平及其表达的影响。结果显示,Syk基因启动子甲基化水平与鼻咽癌细胞分化程度呈负相关,两种鼻咽癌细胞株的Syk mRNA和蛋白质表达水平显著低于NP-69细胞(P<0.01);经5-aza-CdR处理后两种鼻咽癌细胞株的Syk基因启动子甲基化水平降低,Syk mRNA及蛋白质表达升高(P<0.05);高分化鼻咽癌细胞株对药物敏感性高于低分化鼻咽癌细胞株(P<0.01)。由此可见,两种鼻咽癌细胞株中存在不同程度的Syk基因启动子甲基化状态,5-aza-CdR能有效逆转鼻咽癌细胞株Syk基因启动子的甲基化状态,升高Syk mRNA及蛋白质表达,同时鼻咽癌细胞分化程度越高恢复Syk基因表达的比率越高。
The nasopharyngeal carcinoma cell lines CNE-1, CNE-2 and immortalized non-cancerous cells were treated with 5-aza-2’-deoxycytidine (5-aza-CdR) Human nasopharyngeal epithelial cell line NP-69 was used to detect the methylation status of Syk promoter in each cell line before and after 5μmol / L 5-aza-CdR treatment by using the methods of Q-RT-PCR and Western blotting And SykRNA and protein expression. To investigate the methylation of spleen tyrosine kinase (Syk) promoter in 5-aza-2’-deoxycytidine (5-aza-CdR) The impact of its expression. The results showed that the methylation level of Syk gene was negatively correlated with the degree of differentiation of nasopharyngeal carcinoma cells. The expression levels of Syk mRNA and protein in the two nasopharyngeal carcinoma cell lines were significantly lower than those in NP-69 cells (P <0.01) Syk gene promoter methylation levels and Syk mRNA and protein expression were significantly increased in both NPC cell lines after 5-aza-CdR treatment (P <0.05). Highly-differentiated nasopharyngeal carcinoma cell lines were highly sensitive to drugs In poorly differentiated nasopharyngeal carcinoma cell lines (P <0.01). Thus, the methylation status of Syk gene promoter in both nasopharyngeal carcinoma cell lines was different, and 5-aza-CdR could effectively reverse the methylation status of Syk gene promoter in nasopharyngeal carcinoma cell lines Syk mRNA and protein expression, while the higher the degree of nasopharyngeal carcinoma cells to restore Syk gene expression rate.