Methylation and expression of RASSF3 in rat and mouse normal pituitary samples and somatotroph adenoma cell lines.

Methylation levels were investigated using pyrosequencing and are shown as an average level of the 10 CpG sites in the RASSF3 promoter region. RASSF3 mRNA expression fold was determined using qRT-PCR, and β-actin mRNA level was also analyzed as endogenous control. In both GH3 and GT1.1 cells, RASSF3 methylation level was higher and mRNA level was lower than in normal pituitary samples.本研究采用焦磷酸测序（pyrosequencing）检测甲基化水平，结果以RASSF3基因启动子区域内10个CpG位点（CpG sites）的平均甲基化水平进行展示。采用实时定量反转录PCR（qRT-PCR）测定RASSF3的mRNA表达倍数，并以β-肌动蛋白（β-actin）的mRNA水平作为内参对照开展分析。相较于正常垂体样本，GH3与GT1.1两种细胞系中RASSF3的甲基化水平更高，而其mRNA表达水平则更低。