卵泡刺激素剂量依赖性表达基因的筛选

为开发能高效检测卵泡刺激素（follicle-stimulating hormone, FSH）制品生物活性的新方法，选取猪卵泡颗粒细胞，利用转录组测序技术对比FSH处理组（40 mIU/mL）和空白对照组细胞中基因的表达情况，分析并获得大量受FSH调控的候选基因，运用荧光定量聚合酶链式反应验证FSH处理后显著上调的基因，并进一步利用FSH浓度梯度（0、20、40、60、80、100 mIU/mL）处理卵泡颗粒细胞。结果显示，基因OLR1、LHCGR、SCARA5 和S100A12 的表达量呈现FSH浓度依赖性升高。在3 倍递增的FSH浓度梯度（0、1、3、9、27、81、243、729 mIU/mL）下观察各基因表达量的变化，发现在0~243 mIU/mL 的FSH 浓度范围内，LHCGR 基因的表达量与FSH 浓度呈高度线性相关（R2=0.982）。因此，可通过检测FSH刺激条件下细胞中LHCGR基因的表达量，来评估FSH制品的生物学效价。 通讯作者: 石放雄（https://orcid.org/0000-0001-6271-6184）;潘建治（https://orcid.org/0000-0003-3505-0983) E-mail: fxshi@njau.edu.cn; jzpan9@126.com
Abstract: To develop a new and effective method for detection of follicle-stimulating hormone (FSH) bioactivity, porcine follicular granulosa cells were used as experimental materials, and expression of genes in FSH-treated group (40 mIU/mL) and blank control group was compared by transcriptome sequencing to identify FSH-regulated candidate genes. Meanwhile, real-time fluorescent quantitative polymerase chain reaction was used to validate significantly upregulated genes after FSH treatments, and the porcine follicular granulosa cells were treated further by FSH dose gradients (0, 20, 40, 60, 80 and 100 mIU/mL). The results indicated that the expressions of OLR1, LHCGR, SCARA5 and S100A12 increased in a FSH dose-dependent manner. In addition, the expression of these four genes was determined in FSH dose gradients (0, 1, 3, 9, 27, 81, 243 and 729 mIU/mL), and the results showed that the correlation coefficient between the expression of LHCGR and dose gradient of FSH was close to 1 (R2=0.982) in the FSH dose range of 0-243 mIU/mL. In summary, LHCGR expression has a linear relationship with FSH dose, which may be helpful to develop a novel method for biological value detection of FSH. Key words: follicle-stimulating hormone follicular granulosa cell biological value pig