Department of Burn, Wound Healing Center, Shanghai Burn Institute, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
Wound Healing Center, Department of Burn, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
BMC Endocr Disord. 2023 Sep 19;23(1):200. doi: 10.1186/s12902-023-01430-2.
Diabetes foot ulcer (DFU) is a serious complication of diabetes, which can lead to significant mortality and amputation rate. Our previous study found circ_072697 was highly expressed in DFU tissues, but the regulatory mechanism of circ_072697 in DFU remains unclear.
The relative expressions of circ_072697, miR-3150a-3p, and KDM2A in DFU patients or advanced glycation end products (AGEs)-treated HaCaT cells (used as DFU cell model) were determined by using qRT-PCR. Cell proliferation and migration abilities were determined by using CCK-8 and Transwell assays. The interaction between miR-3150a-3p with circ_072697 or KDM2A were verified by RNA immunoprecipitation (RIP) and dual-luciferase reporter assays. Furthermore, the protein expression of genes involved in MAPK signaling pathway was detected by western blot.
The expression of circ_072697 was significantly upregulated in DFU tissues, while the expression of miR-3150a-3p was downregulated. Circ_072697 knockdown promoted the proliferation and migration of AGEs-treated HaCaT cells. miR-3150a-3p was confirmed as a target of circ_072697 and its inhibitor reversed the promotion effects of circ_072697 knockdown on biological behavior of cells. In addition, KDM2A was considered as a target of miR-3150a-3p and it was highly expressed in DFU samples. Importantly, circ_072697 could regulate KDM2A expression through sponging miR-3150a-3p, and this axis had effect on the MAPK signaling pathway.
Overall, circ_072697 regulated the biological behaviors of keratinocytes in DFU via miR-3150a-3p/KDM2A axis and MAPK signaling pathway, revealing a new insight into the pathogenesis and potential therapeutic targets of DFU.
糖尿病足溃疡（DFU）是糖尿病的一种严重并发症，可导致较高的死亡率和截肢率。我们之前的研究发现，circ_072697 在 DFU 组织中高表达，但 circ_072697 在 DFU 中的调控机制尚不清楚。
采用 qRT-PCR 检测 DFU 患者或晚期糖基化终产物（AGEs）处理的 HaCaT 细胞（用作 DFU 细胞模型）中 circ_072697、miR-3150a-3p 和 KDM2A 的相对表达。通过 CCK-8 和 Transwell 实验测定细胞增殖和迁移能力。采用 RNA 免疫沉淀（RIP）和双荧光素酶报告实验验证 miR-3150a-3p 与 circ_072697 或 KDM2A 的相互作用。此外，通过 Western blot 检测 MAPK 信号通路相关基因的蛋白表达。
DFU 组织中 circ_072697 的表达明显上调，而 miR-3150a-3p 的表达下调。circ_072697 敲低促进了 AGEs 处理的 HaCaT 细胞的增殖和迁移。miR-3150a-3p 被证实为 circ_072697 的靶基因，其抑制剂逆转了 circ_072697 敲低对细胞生物学行为的促进作用。此外，KDM2A 被认为是 miR-3150a-3p 的靶基因，在 DFU 样本中高表达。重要的是，circ_072697 通过海绵吸附 miR-3150a-3p 来调节 KDM2A 的表达，该轴对 MAPK 信号通路有影响。
总之，circ_072697 通过 miR-3150a-3p/KDM2A 轴和 MAPK 信号通路调节 DFU 角质形成细胞的生物学行为，为 DFU 的发病机制和潜在治疗靶点提供了新的见解。
