CircST6GAL1 knockdown alleviates pulmonary arterial hypertension by regulating miR-509-5p/multiple C2 and
Xing Zhang1, Hao Qin1, Qiang Ma1
1Department of Peripheral Vascular Diseases, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Insights
Circular RNA circST6GAL1 promotes pulmonary arterial hypertension (PAH) by enhancing cell proliferation and migration. Silencing circST6GAL1 alleviates PAH through the miR-509-5p/MCTP2 pathway, offering a potential therapeutic target.
Area of Science:
- Molecular Biology
- Cardiovascular Research
- RNA Biology
Background:
- Hypertension is a major risk factor for cardiovascular diseases.
- Deregulation of circular RNAs (circRNAs) plays a role in the pathogenesis of pulmonary arterial hypertension (PAH).
- This study investigates the role and mechanism of circST6GAL1 in PAH.
Purpose of the Study:
- To evaluate the function and mechanism of circST6GAL1 in the context of pulmonary arterial hypertension (PAH).
- To determine the molecular interactions involving circST6GAL1, miR-509-5p, and MCTP2 in PAH pathogenesis.
- To assess the therapeutic potential of targeting circST6GAL1 in PAH.
Main Methods:
- Human pulmonary artery smooth muscle cells (HPASMCs) were cultured under hypoxic conditions.
- Cell proliferation, migration, and apoptosis were assessed using CCK-8, EdU, wound healing, and flow cytometry assays.
- Gene and protein expression levels were measured by qRT-PCR and Western blotting; molecular interactions were confirmed via dual-luciferase reporter, RNA immunoprecipitation, and pull-down assays.
- Monocrotaline (MCT)-induced PAH mouse models were used for in vivo validation.
Main Results:
- CircST6GAL1 expression was significantly elevated in PAH patients and hypoxia-induced HPASMCs.
- CircST6GAL1 deficiency inhibited hypoxia-induced HPASMC proliferation and migration while promoting apoptosis.
- CircST6GAL1 directly targets miR-509-5p, which in turn targets MCTP2; circST6GAL1 silencing ameliorated MCT-induced PAH in vivo.
Conclusions:
- CircST6GAL1 deficiency reverses hypoxia-induced cellular changes in HPASMCs and alleviates pulmonary vascular remodeling in a mouse model of PAH.
- The observed effects are mediated through the circST6GAL1/miR-509-5p/MCTP2 axis.
- CircST6GAL1 represents a potential therapeutic target for managing pulmonary arterial hypertension.
Background:
Hypertension is a main contributing factor of cardiovascular diseases; deregulated circular RNAs are involved in the pathogenesis of pulmonary arterial hypertension (PAH). Herein, we evaluated the function and mechanism of circST6GAL1 in PAH process.
Methods:
Human pulmonary artery smooth muscle cells (HPASMCs) were cultured in hypoxic environment for functional analysis. The cell counting kit-8, 5-ethynyl-2'-deoxyuridine, wound healing, and flow cytometry assays were used to investigate cell proliferation, migration, and apoptosis. qRT-PCR and Western blotting analyses were used for level measurement of genes and proteins. The binding between miR-509-5p and circST6GAL1 or multiple C2 and transmembrane domain containing 2 (MCTP2) was analyzed by dual-luciferase reporter, RNA immunoprecipitation, and pull-down assays. The monocrotaline (MCT)-induced PAH mouse models were established for in vivo assay.
Results:
CircST6GAL1 was highly expressed in PAH patients and hypoxia-induced HPASMCs. Functionally, circST6GAL1 deficiency reversed hypoxia-induced proliferation and migration, as well as apoptosis arrest in HPASMCs. Mechanistically, circST6GAL1 directly targeted miR-509-5p, and MCTP2 was a target of miR-509-5p. Rescue assays showed that the regulatory effects of circST6GAL1 deficiency on hypoxia-induced HPASMCs were abolished. Moreover, forced expression of miR-509-5p suppressed HPASMC proliferation and migration and induced cell apoptosis under hypoxia stimulation, while these effects were abolished by MCTP2 overexpression. Moreover, circST6GAL1 silencing improved MCT-induced pulmonary vascular remodeling and PAH.
Conclusion:
CircST6GAL1 deficiency reversed hypoxia-induced proliferation and migration, as well as apoptosis arrest in HPASMCs, and alleviated pulmonary vascular remodeling in MCT-induced PAH mouse models through the miR-509-5p/MCTP2 axis, indicating a potential therapeutic target for PAH.
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