背景:糖尿病性白内障(DC)是糖尿病的常见并发症,其病因和进展是多因素的。在这项研究中,探讨了特异性蛋白1(SP1)和成纤维细胞生长因子7(FGF7)在DC发育中的作用。
方法:用高糖处理SRA01/04细胞建立DC细胞模型。进行MTT测定以评估细胞活力。进行Transwell测定和伤口愈合测定以评估细胞迁移和侵袭。Westernblot和qRT-PCR检测N-cadherin的表达,E-cadherin,胶原蛋白I,纤连蛋白,SP1和FGF7表达。进行CHIP测定和双荧光素酶报告基因测定以分析FGF7和SP1之间的组合。
结果:FGF7在DC患者和HG诱导的SRA01/04细胞中上调。HG处理促进SRA01/04细胞活力,迁移,侵袭和上皮间质转化(EMT),而FGF7敲除减轻了影响。转录因子SP1激活了FGF7的转录水平,SP1过表达加重了HG诱导的SRA01/04细胞损伤。SP1沉默抑制了HG诱导的SRA01/04细胞活力,迁移,入侵和EMT,但这些影响通过上调FGF7得到改善。此外,SP1敲低通过调节FGF7的转录水平抑制PI3K/AKT通路。
结论:转录因子SP1激活FGF7的转录水平和PI3K/AKT通路,调节HG诱导的SRA01/04细胞活力,迁移,入侵和EMT。
BACKGROUND: Diabetic cataract (DC) is a common complication of diabetes and its etiology and progression are multi-factorial. In this study, the roles of specific protein 1 (SP1) and fibroblast growth factor 7 (FGF7) in DC development were explored.
METHODS: DC cell model was established by treating SRA01/04 cells with high glucose (HG). MTT assay was conducted to evaluate cell viability. Transwell assay and wound-healing assay were performed to assess cell migration and invasion. Western blot assay and qRT-PCR assay were conducted to measure the expression of N-cadherin, E-cadherin, Collagen I, Fibronectin, SP1 and FGF7 expression. CHIP assay and dual-luciferase reporter assay were conducted to analyze the combination between FGF7 and SP1.
RESULTS: FGF7 was upregulated in DC patients and HG-induced SRA01/04 cells. HG treatment promoted SRA01/04 cell viability, migration, invasion and epithelial-mesenchymal transition (EMT), while FGF7 knockdown abated the effects. Transcription factor SP1 activated the transcription level of FGF7 and SP1 overexpression aggravated HG-induced SRA01/04 cell injury. SP1 silencing repressed HG-induced SRA01/04 cell viability, migration, invasion and EMT, but these effects were ameliorated by upregulating FGF7. Additionally, SP1 knockdown inhibited the PI3K/AKT pathway by regulating the transcription level of FGF7.
CONCLUSIONS: Transcription factor SP1 activated the transcription level of FGF7 and the PI3K/AKT pathway to regulate HG-induced SRA01/04 cell viability, migration, invasion and EMT.