The MiR-182-5p Antagomir Enhances The Sensitivity Of Gastric Cancer Cell Lines To 5-FU

Objective According to drug resistance of 5-fluorouracil(5-FU)in patients with gastric cancer,we aimed to investigate the effect on the combination of mi R-182-5p antagomir and 5-FU on the proliferation,apoptosis and migration of 5-FU resistance human gastric cancer(SGC7901/5-FU)cell lines and to study the mechanism of mi R-182-5p antagomir increased the sensitization of SGC7901/5-FU cell lines to 5-FU.Methods The SGC7901/5-FU cell lines were selected and divided into negative control(NTC)group,mi R-182-5p antagomir group,5-FU group and mi R-182-5p antagomir in combination with 5-FU group in this study.Firstly,the NTC and mi R-182-5p antagomir were transfected into SGC7901/5-FU cell lines by lipofection,and the expression of mi R-182-5p was detected by quantitative real-time PCR(q RT-PCR).According to grouping,the proliferation was detected by MTT assay in four groups of SGC7901/5-FU cell lines.Flow cytometry was used to detect the apoptosis in four groups of SGC7901/5-FU cell lines and transwell assay was conducted to detect the effect on migration in four groups of SGC7901/5-FU cell lines.q RT-PCR was used to detect the gene expression level of Wnt1,β-catenin,and c-myc in Wnt/β-catenin signaling pathway in four groups of SGC7901/5-FU cell lines.Western blot was used to detect the protein expression of Wnt1,β-catenin and c-myc in Wnt/β-catenin signaling pathway in four groups of SGC7901/5-FU cell lines.Results Compared with NTC group,the expression level of mi R-182-5p was decreased in SGC7901/5-FU cell lines after transfection with mi R-182-5p antagomir(t=18.89,P<0.05).Compared with 5-FU group,mi R-182-5p antagomir group and 5-FU group,the expression level of mi R-182-5p was also decreased in mi R-182-5p antagomir combined with 5-FU group(F=105.22,P<0.05).Compared with 5-FU group,mi R-182-5p antagomir group and 5-FU group,mi R-182-5p antagomir combined with 5-FU group had a decreased cell survial rate and migration rate of SGC7901/5-FU cell lines,and had a increased cell apoptosis rate of SGC7901/5-FU cell lines,and the difference was statistically significant(F=182.5,107.38,490.53,P<0.05).Compared with NTC group,mi R-182-5p antagomir group and 5-FU group,q RT-PCR found that the genes expression level of Wnt1,β-catenin and c-myc were decreased in mi R-182-5p antagomir combined with 5-FU group(F=125.12,82.88,110.20,P<0.05).Western blot indicated that the proteins expression of Wnt1,β-catenin and c-myc were decreased in mi R-182-5p antagomir combined with 5-FU group(F=260.38,53.72,12.74,P<0.05).Conclusion 1.The combination of mi R-182-5p antagomir and 5-FU can significantly down-regulate mi R-182-5p and reduce the proliferation and migration,promote cell apoptosis of SGC7901/5-FU cell lines.mi R-182-5p antagomir increased the sensitization of SGC7901/5-FU cell lines to 5-FU chemotherapy.2.The combination of mi R-182-5p antagomir and 5-FU can significantly down-regulate the expression of Wnt1,β-catenin and c-myc genes and proteins in Wnt/β-catenin signaling pathway.The mechanism may be through negative regulation of Wnt/β-catenin signaling pathway.