Overexpressed lncRNA AC068039.4 Contributes to Proliferation and Cell Cycle Progression of Pulmonary Artery Smooth Muscle Cells Via Sponging miR-26a-5p/TRPC6 in Hypoxic Pulmonary Arterial Hypertension.

Yuhan Qin,Linqing Li,Erfei Luo,Bo Liu,Gaoliang Yan,Chengchun Tang,Dong Wang,Boqian Zhu,Yong Qiao,Jiantong Hou

doi:10.1097/shk.0000000000001606

Abstract

Hypoxic pulmonary hypertension (HPH) is a devastating and incurable disease characterized by pulmonary vascular remodeling, resulting in right heart failure and even death. Accumulated evidence has confirmed long coding RNAs (lncRNAs) are involved in hypoxia-induced pulmonary vascular remodeling in HPH. The exact mechanism of lncRNA in hypoxic pulmonary hypertension remains unclear. Microarray analysis was applied to investigate the profiles of lncRNA expression in pulmonary artery smooth muscle cells (PASMCs) cultured under hypoxia and normoxia condition. qRT-PCR was performed for the expression of lncRNAs, miRNA, and mRNAs, western blot analysis was employed for the detection of the expression of proteins. CCK-8 and transwell chamber assay were applied for the assessment of PASMC proliferation and migration, respectively. Besides, flow cytometry was performed for assessments of cell cycle progression. The binding between AC068039.4 and miR-26a-5p, miR-26a-5p, and TRPC6 3'UTR was detected by dual luciferase reporter assay. A total of 1,211 lncRNAs (698 up-regulated and 513 down-regulated) were differently expressed in hypoxia-induced PASMCs. Consistent with microarray analysis, quantitative PCR verified that AC068039.4 was obviously up-regulated in hypoxia-induced PASMCs. Knocking down AC068039.4 alleviated proliferation and migration of PASMCs and regulated cell cycle progression through inhibiting cells entering the G0/G1 cell cycle phase. Further experiment indicated AC068039.4 promoted hypoxic PASMCs proliferation via sponging miR-26-5p. In addition, transient receptor potential canonical 6 (TRPC6) was confirmed to be a target gene of miR-26a-5p. In conclusion, downregulation of lncRNA AC068039.4 inhibited pulmonary vascular remodeling through AC068039.4/miR-26a-5p/TRPC6 axis, providing new therapeutic insights for the treatment of HPH.

Highlights

Pulmonary hypertension (PH) is an incurable disease with complex pathogenesis and characterized by over-proliferation, apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs), pulmonary vascular remodeling (PVR) and elevated pulmonary vascular resistance, resulting in increased pulmonary arterial pressure, right heart failure and even death[1, 2]
Hierarchical cluster analysis showed that the expression profiles of long coding RNAs (lncRNAs) in PASMCs induced by normoxia and hypoxia were different (Fig. 1A)
Compared with the control group, a total of 1211 lncRNAs in PASMCs induced by hypoxia were significantly differently expressed (698 up-regulated and 513 down-regulated lncRNAs), indicating that these lncRNAs may be involved in the development of hypoxic pulmonary hypertension

Summary

Introduction

Pulmonary hypertension (PH) is an incurable disease with complex pathogenesis and characterized by over-proliferation, apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs), pulmonary vascular remodeling (PVR) and elevated pulmonary vascular resistance, resulting in increased pulmonary arterial pressure, right heart failure and even death[1, 2]. Clinical symptoms could be relieved under current treatments, no effective treatments for HPH have been developed and disease progression remains inevitable[5]. Hypoxic pulmonary hypertension (HPH) is a devastating and incurable disease characterized by pulmonary vascular remodeling, resulting to right heart failure and even death. Accumulated evidence has confirmed long coding RNAs (lncRNAs) are involved in hypoxia induced pulmonary vascular remodeling in HPH. The exact mechanism of lncRNA in hypoxic pulmonary hypertension remains unclear

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Discussion

Conclusion