摘要
目的检测胶质瘤组织和细胞系中Nanog基因启动子区甲基化状态与Nanog基因的表达,并探讨其在胶质瘤发展中的作用。方法采用甲基化特异性PCR(MSP)法分别检测正常脑组织、胶质瘤组织、胶质瘤癌旁组织、胶质瘤细胞系U87和U251中Nanog基因启动子区甲基化状态,实时定量PCR(RT-PCR)法检测相应组织和细胞系中Nanog表达情况。结果 MSP法检测胶质瘤组织Nanog基因启动子区呈非甲基化状态,且非甲基化检出率(58.82%)高于癌旁组织(13.63%),差异有统计学意义(χ2=14.852,P<0.01)。对MSP法检测结果行灰度值分析,高级别胶质瘤中Nanog基因启动子区非甲基化程度高于低级别组。U87和U251细胞系中Nanog基因启动子区呈非甲基化状态。RT-PCR结果显示高级别胶质瘤中Nanog mRNA相对表达量高于低级别组,U87和U251细胞系中Nanog mRNA的转录明显增多。结论胶质瘤中Nanog基因启动子区呈非甲基化状态,且可能促进Nanog基因的转录,影响胶质瘤的发生与发展。
Objective To detect the methylation status of Nanog gene promoter and the expression of Nanog mRNA in gliomas and glioma cell lines, and to explore the function of Nanog in glioma. Methods MSP and RT-PCR were conducted to detect Nanog gene promoter′s methylation status and its mRNA expression in normal brain tissues, gli-oma tissues, paracancerous tissues, glioma cell lines U87 and U251. Results Nanog gene promoter was hyperm-ethylated in gliomas, and the presence was significantly higher in gliomas than that in the paracancerous tissues （58. 82% vs 13. 63% ）（χ2 = 14. 852,P 〈 0. 01）. Nanog gene promoter unmethylation state was higher in grade Ⅲand grade Ⅳ glioma compared with grade I and grade Ⅱ tumors. Nanog gene promoter was hypermethylated in U87 and U251 cell lines. Nanog mRNA expression levels in high-grade（Ⅲ ＋ Ⅳ） gliomas were higher than the low-grade（I ＋ Ⅱ） group, Nanog mRNA expression was significantly increased in U87 and U251 cell lines. Conclusion Unmethylation state of Nanog gene promoter regions is one of the most important mechanisms that up-regulate its protein expression and influence the occurrence and development of gliomas.
出处
《安徽医科大学学报》
CAS
北大核心
2015年第8期1068-1071,共4页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81172407)
安徽省重点实验室绩效考核项目(编号:1306c083028)
