Department of Hepatic biliary Pancreatic Medicine, First Hospital of Jilin University, Changchun, China.
Department of Hepatic biliary Pancreatic Medicine, First Hospital of Jilin University, Changchun, China; Department of Infectious Diseases, First Hospital of Jilin University, Changchun, China.
Toxicol Appl Pharmacol. 2024 Oct;491:117078. doi: 10.1016/j.taap.2024.117078. Epub 2024 Aug 28.
RUNX1 with CBFβ functions as an activator or repressor of critical mediators regulating cellular function. The aims of this study were to clarify the role of RUNX1 on regulating TGF-β1-induced COL1 synthesis and the mechanism of calcipotriol (Cal) on antagonizing COL1 synthesis in PSCs. RT-qPCR and Western Blot for determining the mRNAs and proteins of RUNX1 and COL1A1/1A2 in rat PSC line (RP-2 cell). Luciferase activities driven by RUNX1 or COL1A1 or COL1A2 promoter, co-immunoprecipitation and immunoblotting for pSmad3/RUNX1 or CBFβ/RUNX1, and knockdown or upregulation of Smad3 and RUNX1 were used. RUNX1 production was regulated by TGF-β1/pSmad3 signaling pathway in RP-2 cells. RUNX1 formed a coactivator with CBFβ in TGF-β1-treated RP-2 cells to regulate the transcriptions of COL1A1/1A2 mRNAs under a fashion of pSmad3/RUNX1/CBFβ complex. However, Cal effectively abrogated the levels of COL1A1/1A2 transcripts in TGF-β1-treated RP-2 cells by downregulating RUNX1 production and hindering the formation of pSmad3/RUNX1/CBFβ complexes. This study suggests that RUNX1 may be a promising antifibrotic target for the treatment of chronic pancreatitis.
RUNX1 与 CBFβ 共同作用,作为调节细胞功能的关键介质的激活剂或抑制剂。本研究旨在阐明 RUNX1 在调节 TGF-β1 诱导的 COL1 合成中的作用,以及钙泊三醇(Cal)拮抗 PSCs 中 COL1 合成的机制。通过 RT-qPCR 和 Western Blot 检测大鼠 PSC 系(RP-2 细胞)中 RUNX1 和 COL1A1/1A2 的 mRNAs 和蛋白。使用 RUNX1 或 COL1A1 或 COL1A2 启动子驱动的荧光素酶活性、共免疫沉淀和免疫印迹分析 pSmad3/RUNX1 或 CBFβ/RUNX1,以及 Smad3 和 RUNX1 的敲低或上调。TGF-β1/pSmad3 信号通路调节 RP-2 细胞中 RUNX1 的产生。RUNX1 在 TGF-β1 处理的 RP-2 细胞中与 CBFβ 形成共激活子,以调节 COL1A1/1A2 mRNAs 的转录,方式为 pSmad3/RUNX1/CBFβ 复合物。然而,Cal 通过下调 RUNX1 的产生并阻碍 pSmad3/RUNX1/CBFβ 复合物的形成,有效降低 TGF-β1 处理的 RP-2 细胞中 COL1A1/1A2 转录本的水平。本研究表明,RUNX1 可能是治疗慢性胰腺炎的一种有前途的抗纤维化靶点。