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
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
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DOI：
10.21203/rs.3.rs-25931/v1
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发表时间：
2020-05
影响因子：
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通讯作者：
Yuhan Qin;Boqian Zhu;Linqing Li;G. Yan;Dong Wang;Yong Qiao;Bo Liu;E. Luo;J. Hou;Chengchun Tang
中科院分区：
文献类型：
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作者：
Yuhan Qin;Boqian Zhu;Linqing Li;G. Yan;Dong Wang;Yong Qiao;Bo Liu;E. Luo;J. Hou;Chengchun Tang
背景资料：低血压性肺动脉高压（HPH）是一种以肺血管重构为特征的严重疾病，可导致右心衰竭甚至死亡。大量证据证实长链编码RNA（lncRNA）参与了HPH缺氧诱导的肺血管重构。lncRNA在低氧性肺动脉高压中的确切作用机制尚不清楚。方法：应用基因芯片技术，观察缺氧和常氧条件下培养的肺动脉平滑肌细胞（PASMCs）lncRNA表达的变化。采用qRT-PCR检测lncRNA、miRNA和mRNA的表达，western blot检测蛋白质的表达。CCK-8法检测PASMC增殖，transwell小室法检测PASMC迁移。此外，还进行流式细胞术评估细胞周期进程。结果：低氧诱导的PASMCs共表达1211条lncRNA，其中上调表达698条，下调表达513条。与基因芯片分析结果一致，定量PCR证实AC068039.4在低氧诱导的PASMCs中表达明显上调。敲低AC068039.4可抑制PASMCs的增殖和迁移，并通过抑制细胞进入G0/G1期来调节细胞周期进程。进一步的实验表明，AC068039.4可通过吸收miR-26 - 5p促进缺氧PASMCs的增殖。此外，瞬时受体电位典范6（TRPC 6）被证实是miR-26a-5p的靶基因。结论：总之，下调lncRNA AC068039.4通过AC068039.4/miR-26a-5p/TRPC 6轴抑制肺血管重构，为HPH的治疗提供了新的治疗思路。
Background: 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. Methods：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 detection the expression of proteins. CCK-8 and transwell chamber assay were applied for 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 were detected by dual luciferase reporter assay.Results：A total of 1211 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. Conclusion: 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.