Dongni Wang1, Haiyan Wang1, Cun Liu1, Xiaofeng Mu2, Shaoyun Cheng3. 1. Department of Clinical Laboratory, The 3rd People's Hospital of Qingdao, Qingdao 266041, Shandong Province, China. 2. Department of Clinical Laboratory, Qingdao Central Hospital, Qingdao 266042, Shandong Province, China. 3. Department of Clinical Laboratory, The 3rd People's Hospital of Qingdao, Qingdao 266041, Shandong Province, China. Electronic address: chengshaoyun@126.com.
Abstract
OBJECTIVE: MicroRNAs (miRNAs) have emerged as promising regulators of diabetes mellitus (DM)-induced angiogenic dysfunction in endothelial cells (ECs), but information vis-à-vis the functional roles of distinct miRNAs remain surprisingly scarce. The current study was designed to elucidate the expression and function of miR-140-3p in diabetic ECs. METHODS: miR-140-3p expression was evaluated in DM mouse model and in human ECs using RT-qPCR, Northern blot and RNA fluorescent in situ hybridization. Effects of miR-140-3p manipulation on ECs function were evaluated using cell proliferation, migration and in vitro tube formation assay. Regulation of FOXK2 transcription by miR-140-3p was determined by luciferase reporter assay and site-directed mutagenesis. RESULTS: miR-140-3p expression was significantly down-regulated in high glucose-challenged ECs. Under normal conditions, miR-140-3p knockdown impaired endothelial proliferation and migration, and endothelial tube formation. Mechanistically, miR-140-3p exhibited its proangiogenic effects through directly inhibiting the expression of the forkhead transcription factor FOXK2. From a therapeutic standpoint, shRNA-mediated stable inhibition of FOXK2 effectively corrected miR-140-3p deficiency-induced impairment of ECs proliferation and in vitro angiogenesis. CONCLUSION: Endothelial miR-140-3p positive regulates ECs function by directly targeting FOXK2 signaling. Deregulation of miR-140-3p/FOXK2 cascade by hyperglycemia thus serves as an important contributor to angiogenic dysfunction in DM.
OBJECTIVE: MicroRNAs (miRNAs) have emerged as promising regulators of diabetes mellitus (DM)-induced angiogenic dysfunction in endothelial cells (ECs), but information vis-à-vis the functional roles of distinct miRNAs remain surprisingly scarce. The current study was designed to elucidate the expression and function of miR-140-3p in diabetic ECs. METHODS:miR-140-3p expression was evaluated in DMmouse model and in human ECs using RT-qPCR, Northern blot and RNA fluorescent in situ hybridization. Effects of miR-140-3p manipulation on ECs function were evaluated using cell proliferation, migration and in vitro tube formation assay. Regulation of FOXK2 transcription by miR-140-3p was determined by luciferase reporter assay and site-directed mutagenesis. RESULTS:miR-140-3p expression was significantly down-regulated in high glucose-challenged ECs. Under normal conditions, miR-140-3p knockdown impaired endothelial proliferation and migration, and endothelial tube formation. Mechanistically, miR-140-3p exhibited its proangiogenic effects through directly inhibiting the expression of the forkhead transcription factor FOXK2. From a therapeutic standpoint, shRNA-mediated stable inhibition of FOXK2 effectively corrected miR-140-3pdeficiency-induced impairment of ECs proliferation and in vitro angiogenesis. CONCLUSION: Endothelial miR-140-3p positive regulates ECs function by directly targeting FOXK2 signaling. Deregulation of miR-140-3p/FOXK2 cascade by hyperglycemia thus serves as an important contributor to angiogenic dysfunction in DM.