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目的筛选胶质瘤组织中表达稳定性高的内参基因,为胶质瘤基因表达研究选取合适的内参基因提供实验基础。方法以相对正常脑组织及胶质瘤Ⅰ、Ⅱ、Ⅲ、Ⅳ期组织共25例为实验对象,利用Real-timePCR技术对15个候选内参基因进行筛选,通过geNorm软件对实验结果进行数据分析。结果在15个内参基因中,GUSB、ACTB、TBP、GAPDH、ATP5F1和PPIA6个基因表达稳定,平均表达稳定度(M值)均<0.5,其中ATP5F1和PPIA为表达最稳定的两个内参基因,其M值为0.26。标准化因子V2/3=0.102,小于取舍值0.15。结论在胶质瘤组织实时定量PCR分析中,ATP5F1和PPIA2个内参基因联合使用,可较好地共同校正基因表达结果。
Objective To screen endogenous reference genes with high expression stability in glioma tissues and provide experimental basis for selecting suitable reference genes for the study of gene expression in gliomas. Methods Totally 25 cases of normal brain tissues and gliomas Ⅰ, Ⅱ, Ⅲ and Ⅳ were selected as experimental subjects. Fifteen candidate reference genes were screened by Real-time PCR and the data were analyzed by geNorm software. Results Among the 15 internal reference genes, the expression of GUSB, ACTB, TBP, GAPDH, ATP5F1 and PPIA genes were stable and the average expression stability (M value) were all less than 0.5. ATP5F1 and PPIA were the most stable two internal control genes, The M value is 0.26. Standardization factor V2 / 3 = 0.102, less than the choice of 0.15. Conclusions In real-time quantitative PCR analysis of glioma tissue, two internal reference genes, ATP5F1 and PPIA, can be jointly used to correct the gene expression results.