Experimental Study
Effect of microRNA-384 targeted regulation of Smad nuclear interacting protein 1 on invasion and migration of lung cancer SPC-A1 cells
Dengyan Zhu, Yang Yu, Yang Yang, Donglei Liu, Kai Wu, Yu Qi, Song Zhao
Published 2019-06-08
Cite as Chin J Exp Surg, 2019, 36(6): 1069-1072. DOI: 10.3760/cma.j.issn.1001-9030.2019.06.027
Abstract
ObjectiveTo study the effect of microRNA (miRNA, miR)-384 on invasion and migration of lung cancer SPC-A1 cells and the mechanism of targeted regulation.
MethodsThe miR-384 mimics were transfected into SPC-A1 cells. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to determine the up-regulation effect. The proliferation was measured by methyl thiazol tetrazolium (MTT). Transwell chamber was used to detect invasion and migration changes. Western blotting was used to detect the expression of matrix metalloproteinase (MMP)-2, Vimentin, MMP-9 and E-cadherin. Target gene prediction software predicts that Smad nuclear interacting protein 1 (SNIP1) may be the target gene of miR-384. Luciferase reporting system was used to identify the targeting relationship. The miR-384 mimics and pcDNA 3.1-SNIP1 were co-transfected into SPC-A1 cells. MTT, Transwell chamber and Western blotting were used to detect the effects of pcDNA 3.1-SNIP1 on the proliferation, invasion and migration of up-regulated miR-384 cells.Comparison two groups was performed using t test, and comparison three groups was performed with one-way analysis of variance (ANOVA).
ResultsThe miR-384 mimics could significantly reduce the expression of microRNA-384 in SPC-A1 cells (0.97±0.13 vs. 2.28±0.27), and decrease the cell proliferation (0.41±0.04 vs. 0.29±0.03), invasion (148.23±13.98 vs. 115.30±10.23) and migration (196.48±17.36 vs. 137.58±12.07) ability, inhibit the expression of MMP-2, Vimentin and MMP-9 in cells, and promote the expression of E-cadherin in cells. Luciferase reporter vectors showed that after miR-384 Mimics transfection, the protein level of SNIP1 was significantly decreased (t=9.954, P<0.01), which suggested the expression of SNIP1 was specifically regulated by miR-384. The pcDNA 3.1-SNIP1 could significantly reverse the inhibitory effect of miR-384 mimics on proliferation (0.26±0.04 vs. 0.38±0.04), migration (130.20±10.56 vs. 158.25±11.47) and invasion (110.27±9.68 vs. 149.20±10.38) of SPC-A1 cells. The expression levels of SNIP1, MMP-2, Vimentin and MMP-9 were increased, and the expression levels of E-cadherin were decreased.
ConclusionThe miR-384 targets SNIP1 to inhibit the invasion and migration of lung cancer SPC-A1 cells.
Key words:
Lung cancer; Invasion; MicroRNA-384; Smad nuclear interacting protein 1
Contributor Information
Dengyan Zhu
Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
Yang Yu
Department of Anesthesiology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
Yang Yang
Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
Donglei Liu
Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
Kai Wu
Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
Yu Qi
Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
Song Zhao
Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
