Zhang Xiao-Han, Li Jia-Xin, Wu Xiao-Xu, Zhang Qian, Tian Ming, Yang Si-Qi, Liu Di, Yang Xiu-Qin
College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
Institute of Animal Husbandry, Heilongjiang Academy of Agricultural Sciences, Harbin, 150086, China.
Anim Biosci. 2025 Jan;38(1):41-53. doi: 10.5713/ab.24.0072. Epub 2024 Aug 26.
This study was conducted to reveal the role of nuclear poly(A) binding protein 1 (PABPN1) in the proliferation of preadipocytes, and to reveal the relationship between PABPN1 and cAMP response element (CRE)-binding protein (CREB) in the regulation of preadipocyte proliferation.
Vectors overexpressing and siRNAs against PABPN1/CREB were transiently transfected into both porcine preadipocytes and mouse 3T3-L1 cells. Preadipocyte proliferation was measured with cell counting kit-8, 5-ethynyl-2'-deoxyuridine, real-time quantitative polymerase chain reaction, Western blotting, and flow cytometry analyses. Additionally, the transcriptional regulation of CREB on PABPN1 were analyzed with dual-luciferase reporter gene and electrophoretic mobility shift assay.
Overexpression of PABPN1 inhibits, and knockdown of PABPN1 promotes, the proliferation of both porcine preadipocytes and 3T3-L1 cell lines. PABPN1 overexpression increased, while knockdown decreased, the cell population in the G0/G1 phase. These indicates that PABPN1 repressed preadipocyte proliferation by inhibiting cell cycle progress. Additionally, it was revealed that CREB regulated the expression of PABPN1 through binding to the promoter and that CREB inhibited preadipocyte proliferation by repressed cell cycle progress. Furthermore, we showed that PABPN1 functions as a downstream gene of CREB to regulate the proliferation of preadipocytes.
PABPN1 inhibits preadipocyte proliferation by suppressing the cell cycle. We also found that CREB could promote PABPN1 expression by binding to a motif in the promoter. Further analysis confirmed that PABPN1 functions as a downstream gene of CREB to regulate the proliferation of preadipocytes. These results suggest that the CREB/PABPN1 axis plays a role in the regulation of preadipocyte proliferation, which will contribute to further revealing the mechanism of fat accumulation.
本研究旨在揭示细胞核聚腺苷酸结合蛋白1（PABPN1）在前体脂肪细胞增殖中的作用，并揭示PABPN1与环磷酸腺苷反应元件（CRE）结合蛋白（CREB）在调节前体脂肪细胞增殖中的关系。
将过表达和针对PABPN1/CREB的小干扰RNA（siRNA）载体分别瞬时转染到猪前体脂肪细胞和小鼠3T3-L1细胞中。采用细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷、实时定量聚合酶链反应、蛋白质免疫印迹和流式细胞术分析等方法检测前体脂肪细胞的增殖情况。此外，通过双荧光素酶报告基因和电泳迁移率变动分析，分析CREB对PABPN1的转录调控作用。
PABPN1过表达抑制猪前体脂肪细胞和3T3-L1细胞系的增殖，而敲低PABPN1则促进其增殖。PABPN1过表达使处于G0/G1期的细胞数量增加，而敲低则使其减少。这表明PABPN1通过抑制细胞周期进程来抑制前体脂肪细胞增殖。此外，研究发现CREB通过与启动子结合来调节PABPN1的表达，且CREB通过抑制细胞周期进程来抑制前体脂肪细胞增殖。此外，我们还表明PABPN1作为CREB的下游基因发挥作用，以调节前体脂肪细胞的增殖。
PABPN1通过抑制细胞周期来抑制前体脂肪细胞增殖。我们还发现CREB可通过与启动子中的基序结合来促进PABPN1表达。进一步分析证实，PABPN1作为CREB的下游基因发挥作用，以调节前体脂肪细胞的增殖。这些结果表明，CREB/PABPN1轴在调节前体脂肪细胞增殖中发挥作用，这将有助于进一步揭示脂肪积累的机制。
