Abstract：
Tumour necrosis factor (TNF) superfamily member 11 (TNFSF11), also known as RANKL, plays a crucial role in regulating several physiological and pathological activities. Additionally, it is a vital factor in bone physiology, and the sex hormone progesterone regulates the expansion of stem cells and the proliferation of mammary epithelial cells. It is essential for animal growth and reproductive physiological processes. This study aimed to evaluate the tissue-specific expression characteristics and promoter activity of the TNFSF11 gene in pigs. As a result, the study examined the presence of TNFSF11 expression in the tissues of Xiangsu pigs at 0.6 and 12 months of age. Moreover, the core promoter region of TNFSF11 was also identified by utilizing a combination of bioinformatic prediction and dual-luciferase activity tests. Finally, the effect of transcription factors on the transcriptional activity of the core promoter region was determined using site-directed mutagenesis. TNFSF11 was uniformly expressed in all tissues; however, its expression in muscles was comparatively low. The core promoter region of TNFSF11 was located in the -555 to -1 region. The prediction of the transcription start site of TNFSF11 gene-2000 ∼ + 500bp showed that there was a CpG site in 17 ∼ + 487bp. Analysis of mutations in the transcription factor binding sites revealed that mutations in the Stat5b, Myog, Trl, and EN1 binding sites had significant effects on the transcriptional activity of the TNFSF11 gene, particularly following the EN1 binding site mutation (P < 0.001). This study provides insights into both the tissue-specific expression patterns of TNFSF11 in the tissues of Xiangsu pigs and the potential regulatory effects of transcription factors on its promoter activity. These results may be helpful for future research aimed at clarifying the expression and role of the porcine TNFSF11 gene.
摘要：
肿瘤坏死因子（TNF）超家族成员11（TNFSF11），也被称为RANKL,在调节多种生理和病理活动中起着至关重要的作用。此外,它是骨骼生理学的一个重要因素，性激素孕激素调节干细胞的扩增和乳腺上皮细胞的增殖。它对动物生长和生殖生理过程至关重要。本研究旨在评估猪TNFSF11基因的组织特异性表达特征和启动子活性。因此,该研究检测了0.6和12月龄湘苏猪组织中TNFSF11表达的存在。此外，通过结合生物信息学预测和双荧光素酶活性测试,还鉴定了TNFSF11的核心启动子区.最后,使用定点诱变确定转录因子对核心启动子区转录活性的影响。TNFSF11在所有组织中均匀表达;然而，它在肌肉中的表达相对较低。TNFSF11的核心启动子区位于-555～-1区。对TNFSF11基因转录起始位点-2000~+500bp的预测表明，在17~+487bp中有一个CpG位点。转录因子结合位点的突变分析显示Stat5b中的突变,Myog,Trl,和EN1结合位点对TNFSF11基因的转录活性有显著影响,特别是在EN1结合位点突变后(P<0.001)。这项研究提供了TNFSF11在湘苏猪组织中的组织特异性表达模式以及转录因子对其启动子活性的潜在调节作用的见解。这些结果可能有助于未来旨在阐明猪TNFSF11基因的表达和作用的研究。
