Effect of interleukin-17 in the migration and invasion of gastric cancer cell via regulating epithelial-mesenchymal transition / 中华消化外科杂志

ABSTRACT

Objective To investigate the effect of interleukin-17 (IL-17) in the gastric cancer cell migration and invasion via regulating epithelial-mesenchymal transition (EMT) and its potential function.Methods (1) Human gastric cancer cell MGC-803 lines in the logarithmic growth phase were stimulated by 0, 1 ng/mL, 10 ng/mL,100 ng/mL and 1μg/mL of IL-17 for 48 hours, and the phenotypic changes were observed.The concentration of IL-17 was selected for follow-on experiments based on the most obvious phenotypic changes.Gastric cancer cell MGC-803 which were stimulated by 100 ng/mL of IL-17 and PBS for 48 hours were allocated into the experimental group and control group, respectively.(2) The expressions of E-cadherin and Vimentin mRNA in gastric cancer cells were assayed through real-time PCR (RT-PCR).(3) The relative expressions of E-cadherin and Vimentin proteins in gastric cancer cells were assayed by the Western blot.(4) The scratch test and Transwell detection were also utilized to study the migration and invasion of gastric cancer cell MGC-803 in vitro.Measurement data with normal distribution were presented as-x ± s and comparison between groups was analyzed using the t test.Results (1) There were significant phenotypic changes in the gastric cancer cell after the different concentration of IL-17 stimulated gastric cancer cell MGC-803 for 48 hours.Cells were changed from polygonal and tight junction to spindle and loosely junction with a deterioration of cell adhesion.Cell phenotypes were gradually changed as the concentration of IL-17 was changed from 0 to 100 ng/mL.Phenotypic changes were the most obvious when 100 ng/mL of IL-17 was used, but these were non-significant as the concentration of IL-17 increased to 1 μg/mL with the death and floating of some cells.(2) The relative expressions of E-cadherin mRNA and Vimentin mRNA in RT-PCR were 0.45 ±0.13 and 1.06 ±0.23 in the experimental group and 2.39 ±0.55 and 1.23±0.41 in the control group, respectively, with significant differences (t =3.811, 2.923, P ＜0.05).(3) The results of Western blot showed the relative expressions of E-cadherin and Vimentin proteins were 0.86 ± 0.17 and 1.56 ± 0.29 in the experimental group and 1.01 ± 0.12 and 0.56 ± 0.17 in the control group, respectively, with significant differences (t =3.551, 3.601, P ＜ 0.05).(4) Cell migration in the 2 groups were detected by the scratch test at 36 hours after scratch test, and the width of scratch in the experimental and control groups were (0.76 ± 0.13) mm and (0.40 ± 0.15) mm, showing a significant difference (t =3.095, P ＜ 0.05).Transwell detection showed number of transmembrane cell in the experimental and control groups were 159 ±28 and 94 ± 18, respectively, with a significant difference (t =3.307, P ＜ 0.05).Conclusion IL-17 can promote the migration and invasion of gastric cancer cells via stimulating alteration of EMT.