Effect of small interference RNA silenced S100A4 protein in human gastric carcinoma cells on proliferation, apoptosis and chemotherapy sensitivity in vitro

Abstract

Objective To study the application of small interfering RNA silencing S100A4 protein in human gastric cancer cell BGC-823 proliferation, apoptosis and the effect of chemotherapy sensitivity. Methods Human gastric carcinoma cell line BGC - 823 transfection siRNA, RT-PCR detected the changes of mRNA after transfection. Groups divided into interference group, negative control group and normal control group. MTT test determined different concentrations of oxaliplatin in gastric cancer cells and calculated IC50,then draw cell growth curve, TUNEL method to detect apoptosis, RT-PCR tested each cell mRNA changed, Western blot detected the change of the S100A4 protein. All data analysis by SPSS17.0, t test applied, RT-PCR and Western blot results analysis by SPSS17.0, comparing multiple samples by using single factor analysis of variance and LSD test. P<0.05 was statistically significant. Results RT-PCR results showed that BGC- 823 cell transfection, S100A4mRNA expression quantity respectively after 48 hours: (0.674±0.011), (0.652±0.021), (0.345±0.040), the interference group and normal control group were statistically significant (P=0.012, P 0.380); Western blot results showed BGC - 823 cell transfection S100A4 expression significantly lowered respectively after 48 hours, there were (0.654 ± 0.025), (0.642±0.014), (0.317±0.061), the interference group and normal control group was statistically significant (P=0.01, P 0.341). After S100A4-siRNA transfection, gastric carcinoma BGC-823 cell proliferation decreased, TUNEL method showed obviously increase apoptosis, MTT showed that IC50 of oxaliplatin was 56.31 μmol/L, after transfection, IC50 was 0.654 μmol/L. Conclusions This study showed that the siRNA silence S100A4 protein inhibit gastric cancer cell proliferation, induced apoptosis and improved chemotherapy sensitivity of oxaliplatin. S100A4 might be prompt targets for the treatment of gastric carcinoma. Key words: Gastric carcinoma; siRNA-S100A4; Cell proliferation; Apoptosis; Chemotherapy sensitivity