Cinnamic hydroxamic acid inhibits the proliferation of gastric cancer cells via upregulation of miR 145 expression and down-regulation of P13K/Akt signaling pathway

Shanping Li,Senmao Hu

doi:10.4314/tjpr.v19i5.7

Abstract

Purpose: To investigate the anti-proliferative effect of cinnamic hydroxamic acid (CHA) on gastric cancer (GC) cells, and its mechanism of action.Methods: Two GC cell lines (SGC-7901 and MKN1) and normal human gastric epithelial cells (GES1) were used for this study. The GC cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM)supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin solution at 37 °C for 24 h in a humidified atmosphere of 5 % CO2 and 95 % air. GES1 cells were cultured in RPMI medium supplemented with 10 % FBS only. Cell viability and apoptosis were determined using 3 (4,5 dimethyl thiazol 2 yl) 2,5 diphenyl 2H tetrazolium bromide (MTT), and flow cytometric assays, respectively. The level of expression of microRNA-145 (miR-145) was determined using real-time quantitative polymerase chain reaction (qRT-PCR). Protein expressions of c-Myc, p-AKT, PI3K, p21, and matrix metalloproteinase (MMP)-2 and MMP-9were determined using Western blotting.Results: Treatment of GC cells with CHA for 72 h led to significant and dose-dependent reduction in their viability, and significant and dose-dependent increase in the number of apoptotic cells (p < 0.05). It also significantly arrested GC cell cycle at G1 phase (p < 0.05). The treatment significantly and dosedependently decreased SGC-7901 and MKN1 cell migration and invasion, and upregulated miR-145 mRNA expression (p < 0.05). The expression of miR-145 mRNA was significantly higher in MKN1 cells than in SGC-7901cells (p < 0.05). Treatment of SGC-7901 and MKN1 cells with CHA significantly downregulated protein expressions of c-Myc, MMP-2/9, PI3K and p-AKT, but upregulated p21 protein expression (p< 0.05).Conclusion: These results show that CHA inhibits the proliferation of GC cells via upregulation of miR-145 expression and down-regulation of P13K/Akt signaling pathway. Therefore, CHA has a good potential as a therapeutic agent for the management of gastric cancer Keywords: Apoptosis, Cinnamic hydroxamic acid, Gastric cancer, Metastasis, Proliferation

Highlights

Gastric cancer (GC), a common digestive tract cancer characterized by high morbidity and mortality, is prevalent among South-East Asians [1,2]
This study investigated the antiproliferative effect of Cinnamic hydroxamic acid (CHA) on GC cells, and the mechanism involved
The proliferation of SGC-7901 and MKN1 cells was significantly and dose-dependently suppressed by CHA treatment. These results indicate that CHA may inhibit the proliferation of GC cells via induction of apoptosis and cell cycle arrest at G1 phase

Summary

INTRODUCTION

Gastric cancer (GC), a common digestive tract cancer characterized by high morbidity and mortality, is prevalent among South-East Asians [1,2]. MicroRNAs influence the expression of target genes by pairing with their 3'UTR regions, thereby participating in life processes such as cell growth and differentiation, and apoptosis [6,7]. MiR-145 has a tumor-inhibiting effect, the mechanisms underlying its involvement in the proliferation, metastasis, and invasion of tumor cells are not entirely clear. The. GC cells treated with varied concentrations of CHA (3 – 20 μM) were seeded into 6-well plates and incubated for 48 h. Cells treated with varied concentrations of CHA (3 – 20 μM) for 48 h in RPMI-1640 medium were seeded at a density of 1 x 105 cells/well in Transwell chamber coated with substrate (200 mg/mL Matrigel) and cultured in serum-free medium.

RESULTS

DISCUSSION

Conflict of interest