MicroRNA-30b-5p promotes the proliferation and migration of human airway smooth muscle cells induced by platelet-derived growth factor by targeting phosphatase and tensin homolog deleted on chromosome ten

ABSTRACT

Dysfunction of airway smooth muscle (ASM) cells is crucial in asthma pathogenesis. Here, microRNA-30b-5p (miR-30b-5p)’s function and mechanism in ASM cells’ multiplication and migration were investigated. Microarray was utilized for identifying the differentially expressed miRNAs in the bronchial epithelial cells of the asthma patients and healthy controls. Platelet-derived growth factor (PDGF) was employed to treat ASM cells to establish an in-vitro asthma model. Quantitative real-time PCR (qRT-PCR) was conducted for detecting the expressions of miR-30b-5p and phosphatase and tensin homolog deleted on chromosome 10 (PTEN). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-bromo-2ʹ-deoxyuridine (BrdU) assays were used for examining cell multiplication; Transwell assay was performed for detecting cell migration; cell cycle was analyzed through flow cytometry. The targeted relationship between PTEN and miR-30b-5p was verified using a dual-luciferase reporter gene assay. Western blot was used for detecting the expressions of phosphorylated (p)-phosphatidylinositol 3-kinase (PI3K), PTEN, PI3K, protein kinase B (AKT) and p-AKT in ASM cells. We demonstrated that, miR-30b-5p expression in the bronchial epithelial cells of asthmatic patients was up-regulated. It was also increased in PDGF-stimulated ASM cells. Transfection of miR-30b-5p mimics facilitated ASM cells’ multiplication, migration and cycle progression, while inhibiting miR-30b-5p had the opposite effect. Furthermore, miR-30b-5p could target PTEN to repress PTEN expression. PTEN overexpression attenuated the effect of miR-30b-5p on ASM cells. Moreover, miR-30b-5p overexpression facilitated the expression of p-PI3K and p-AKT in PDGF-stimulated ASM cells. Collectively, miR-30b-5p activates the PI3K/AKT pathway by targeting PTEN to facilitate PDGF-induced dysfunction of ASM cells.

Graphical abstract

KEYWORDS:

• Airway smooth muscle cell
• asthma
• miR-30b-5p
• PTEN
• cell proliferation
• cell migration

Highlights

1. The expression level of miR-30b-5p is reduced in airway smooth muscle cells with platelet-derived growth factor stimulation.

2. MiR-30b-5p represses the growth and migration of airway smooth muscle cells, which contributes to regulating airway remodeling in the pathogenesis of asthma.

3. phosphatase and tensin homolog deleted on chromosome ten is a target gene of miR-30b-5p in airway smooth muscle cells.

List of abbreviations

MiR-30b-5p; microRNA-30b-5p: PTEN; phosphatase and tensin homolog deleted on chromosome ten: ASM, airway smooth muscle; PDGF; platelet-derived growth factor: DMEM; Dulbecco’s modified Eagle’s medium: qRT-PCR; quantitative real-time polymerase chain reaction: TBST; tris buffered saline with tween: PI3K; phosphatidylinositol 3-kinase: AKT; protein kinase B: 3ʹUTR; 3ʹ-untranslated region: MTT; 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide: BrdU; 5-bromo-2ʹ-deoxyuridine: PI; propidium iodide: GEO; gene expression omnibus

Disclosure statement

No potential conflict of interest was reported by the author(s).

Ethics statement

Our study was approved by the Ethics Review Board of Affiliated Hospital of Chengde Medical College.

Data Availability Statement

The data used to support the findings of this study are available from the corresponding author upon request.