Role and mechanism of gastrokine 1 in gastric cancer metastasis

QI Jing-peng; SHI A-ping; FENG Fan; REN Gui; ZHANG Hong-wei

doi:10.3969/j.issn.1002-266X.2015.32.001

Abstract：Objective To investigate the role of gastrokine 1 (GKN1) in the metastasis of gastric cancer and to ob-serve whether it is related with the matrix metalloproteinase (MMP) family and NF-κB pathway.Methods Quantitative PCR was carried out to detect the GKN 1 mRNA expression in the gastric cancer cell lines MGC 803, MKN-45 and non-neo-plastic epithelial cell line HFE-145 of the logarithmic phase .MKN-45 cells in the logarithmic phase were randomly divided into three groups:the transfection group (group A), non-load group (group B) and the control group (group C).Group A was transfected by GKN1 expression vector , group B was transfected by empty vector and the group C was not treated .MTT assay was used to detect the cell proliferation activities of the three groups for 5 days, cell scratch method was used to detect the scratch width, and Western blotting was employed to detect the protein expression of MMP-2, MMP-9 and NF-κB p65. Results The GKN1 mRNA relative expression levels of human gastric cancer cell lines MGC 803 and MKN-45 were 0.172 ±0.024 and 0.231 ±0.031, and the GKN1 relative mRNA expression of the epithelial cells HFE-145 was 1.024 ± 0.264.The GKN1 mRNA expression of the two kinds of human gastric cancer cells was lower than that of HFE -145 cells (all P<0.05).No significant difference was found in the cell proliferation activity among the three groups of MKN -45 cells after being cultured for one or two days (all P>0.05).After being cultured for day 3, 4 and 5, the cell proliferation activ-ity of group A was lower than that of groups B and C (all P<0.05).The scratch width of group A after scratch test for 48 h and 72 h was higher than that of groups B and C (all P<0.05).The expression of MMP-2, MMP-9 and NF-κB p65 of group A was lower than that of groups B and C (all P<0.05), and no significant difference was found between groups B and C (all P>0.05).Conclusions GKN1 in the gastric cancer cell lines shows lower expression and the activation of its expression can inhibit the metastasis of tumor cells , which may be related with its inhibition of MMP protein expression and NF-κB pathway .