Journal of Shanghai Jiao Tong University (Medical Science) >

2022 , Vol. 42 >Issue 11: 1534 - 1541

DOI: https://doi.org/10.3969/j.issn.1674-8115.2022.11.003

Basic research

lncRNA NEAT1 affects the proliferation, invasion and migration of ox-LDL-induced human vascular smooth muscle cells through miR-377-3p/Wnt pathway

  • Yongmei YUAN ,
  • Xiaodan CHENG ,
  • Jiaan SUN ,
  • Dongge CHANG ,
  • Yingying HE ,
  • Chang LIU
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  • Department of Emergency, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou 450007, China
LIU Chang, E-mail: lcjzk0@163.com.

Received date: 2022-04-21

  Accepted date: 2022-08-21

  Online published: 2023-01-04

Supported by

2020 Key Scientific Research Projects of Higher Education Institutions in Henan Province(20B320024)

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Abstract

Objective ·To study the effects of long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) on the proliferation, invasion and migration of oxidized low-density lipoprotein (ox-LDL)-induced human vascular smooth muscle cells (HVSMCs) through regulating microRNA-377-3p (miR-377-3p) /Wnt pathway. Methods ·HVSMCs cultured in vitro were treated with 100 μg/mL ox-LDL. HVSMCs were randomly divided into control group, model group, NEAT1 siRNA group, miR-377-3p inhibitor group, transfection+inhibitor (NEAT1 siRNA+miR-377-3p inhibitor) group, and transfection-negative control (NEAT1 siRNA-negative control) group. Except for the control group, the atherosclerosis cell models were induced with 100 μg/mL ox-LDL in other groups. After grouping and transfection, the cell viability of each group was determined by using cell counting kit-8 (CCK-8) experiment; the apoptosis rate of each group was measured by flow cytometry; the migration and invasion of cells in each group were evaluated by the Transwell experiment; the expression of cell proliferation markers such as proliferating cell nuclear antigen (PCNA), and epithelial-mesenchymal transition (EMT) markers, such as E-cadherin and vimentin in each group, was determined by immunoblotting experiment; the expression of NEAT1, miR-377-3p and wingless-related MMTV integration site 5A (WNT5A) mRNA in each group was determined by qRT-PCR experiment. Results ·Compared with the control group, the mRNA expression of NEAT1 in the model group increased, the mRNA expression of miR-377-3p decreased, the apoptosis rate decreased, the cell viability, and the number of migrating and invasive cells increased, the protein expression of PCNA and Vimentin increased, the expression of E-cadherin decreased, and WNT5A mRNA expression increased (all P=0.000). After NEAT1 was interfered, the apoptosis rate increased, the cell viability, and the number of migrating and invasive cells decreased, the protein expression of PCNA and Vimentin decreased, the expression of E-cadherin increased, and WNT5A mRNA expression decreased (all P=0.000); after miR-377-3p was inhibited, all the above indicators showed opposite expression trends (P<0.05). In addition, inhibition of miR-377-3p reversed the regulation of NEAT1 interference on cell proliferation, invasion, migration and apoptosis (all P=0.000). Conclusion ·The down-regulating of NEAT1 expression may increase miR-377-3p expression by competitive binding to miR-377-3p, reduce protein expression associated with the Wnt pathway, inhibit the abnormal proliferation, invasion and migration of HVSMCs induced by ox-LDL, and induce apoptosis.

Key words： long non-coding RNA (lncRNA); nuclear paraspeckle assembly transcript 1 (NEAT1); microRNA-377-3p (miR-377-3p); human vascular smooth muscle cell (HVSMC); proliferation; invasion and migration

Cite this article

Yongmei YUAN , Xiaodan CHENG , Jiaan SUN , Dongge CHANG , Yingying HE , Chang LIU . lncRNA NEAT1 affects the proliferation, invasion and migration of ox-LDL-induced human vascular smooth muscle cells through miR-377-3p/Wnt pathway[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022 , 42(11) : 1534 -1541 . DOI: 10.3969/j.issn.1674-8115.2022.11.003

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