Wei Liu1, Qimei Gong1, Junqi Ling2, Wen Zhang1, Zhaohui Liu1, Jingjing Quan1. 1. Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China. 2. Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China. Electronic address: lingjq@mail.sysu.edu.cn.
Abstract
INTRODUCTION: Growing evidence shows microRNAs (miRNAs) regulate numerous cellular processes. The purpose of this study was to investigate whether miRNAs can regulate the commitment of human dental pulp cells (hDPCs) to the angiogenic fate. METHODS: The hDPCs were induced to differentiate into the vascular lineage. Gene expression of endothelial markers (vWF and CD31) on day 7 after induction was analyzed by using quantitative reverse transcription-polymerase chain reaction (qRT-PCR).The miRNA expression profiling of endothelial differentiation was performed by microarray and was validated by qRT-PCR analysis. The hDPCs were infected by recombinant lentivirus to overexpress or knock down miR-424 stably, and the biological effects of miR-424 on the endothelial differentiation of hDPCs were further investigated. The tube formation ability and the amount of endothelial markers (vWF and KDR) were evaluated by Matrigel assay and Western blotting. Target genes of miR-424 were further determined by bioinformatic algorithms and Western blotting. RESULTS: After endothelial differentiation, the expression of vWF and CD31 increased significantly in hDPCs. Microarray data showed that the miR-424 expression level was down-regulated on day 7. The qRT-PCR revealed a time-dependent decrease, with significant differences detected on day 1 and day 7 (P < .05). Knockdown of miR-424 expression in hDPCs promoted endothelial differentiation, with increased tube formation and up-regulated expression of vWF and KDR. In contrast, overexpression of miR-424 inhibited their differentiation. In addition, miR-424 was predicted to target vascular endothelial growth factor and KDR. Overexpression of miR-424 decreased vascular endothelial growth factor and KDR protein levels, whereas miR-424 inhibition significantly elevated them. CONCLUSIONS: This study demonstrated that miR-424 may play a negative role in regulating endothelial differentiation of hDPCs, and inhibition of miR-424 may contribute to dental pulp repair and regeneration.
INTRODUCTION: Growing evidence shows microRNAs (miRNAs) regulate numerous cellular processes. The purpose of this study was to investigate whether miRNAs can regulate the commitment of human dental pulp cells (hDPCs) to the angiogenic fate. METHODS: The hDPCs were induced to differentiate into the vascular lineage. Gene expression of endothelial markers (vWF and CD31) on day 7 after induction was analyzed by using quantitative reverse transcription-polymerase chain reaction (qRT-PCR).The miRNA expression profiling of endothelial differentiation was performed by microarray and was validated by qRT-PCR analysis. The hDPCs were infected by recombinant lentivirus to overexpress or knock down miR-424 stably, and the biological effects of miR-424 on the endothelial differentiation of hDPCs were further investigated. The tube formation ability and the amount of endothelial markers (vWF and KDR) were evaluated by Matrigel assay and Western blotting. Target genes of miR-424 were further determined by bioinformatic algorithms and Western blotting. RESULTS: After endothelial differentiation, the expression of vWF and CD31 increased significantly in hDPCs. Microarray data showed that the miR-424 expression level was down-regulated on day 7. The qRT-PCR revealed a time-dependent decrease, with significant differences detected on day 1 and day 7 (P < .05). Knockdown of miR-424 expression in hDPCs promoted endothelial differentiation, with increased tube formation and up-regulated expression of vWF and KDR. In contrast, overexpression of miR-424 inhibited their differentiation. In addition, miR-424 was predicted to target vascular endothelial growth factor and KDR. Overexpression of miR-424 decreased vascular endothelial growth factor and KDR protein levels, whereas miR-424 inhibition significantly elevated them. CONCLUSIONS: This study demonstrated that miR-424 may play a negative role in regulating endothelial differentiation of hDPCs, and inhibition of miR-424 may contribute to dental pulp repair and regeneration.
Authors: Beatriz A Rodas-Junco; Michel Canul-Chan; Rafael A Rojas-Herrera; Clelia De-la-Peña; Geovanny I Nic-Can Journal: Front Physiol Date: 2017-12-06 Impact factor: 4.566
Authors: Ahmed Saad; Xiang-Yang Zhu; Sandra Herrmann; LaTonya Hickson; Hui Tang; Allan B Dietz; Andre J van Wijnen; Lilach Lerman; Stephen Textor Journal: Stem Cell Res Ther Date: 2016-09-09 Impact factor: 6.832