Effect of colon cancer metastasis associated gene-1 on biological function of gastric cancer cell line MGC-803 by regulating hepatocyte growth factor/c-Met signaling pathway

Abstract

Objective To study the effect of colon cancer metastasis associated gene-1 (MACC-1) expression on the proliferation and apoptosis of gastric cancer cell line MGC-803 in vitro. Methods Cultured gastric cancer MGC-803 cells and passaged cells were divided into three groups: group A [MACC-1-short hairpin RNA (shRNA) and PSV2neo3 co transfection group]; group B (shNC and PSV2neo co transfection group); group C (non transfection group). By Western blotting and real-time quantitative polymerase chain reaction (Real-time PCR), the expression of MACC-1 protein and mRNA was detected in the three groups, and that of of c-Met, E-cadherin and N-cadherin in 3 groups of MGC-803 cells were detected. Flow cytometry and scratch test were used to analyze migration, proliferation and apotosis of MGC-803 cells transfected with MACC-1-shRNA. Results (1) Western blotting and Real-time PCR test results showed that as compared with groups B and C, MACC-1 and c-Met protein and mRNA expression in group A were significantly down-regulated (P<0.01). (2) Western blotting and Real-time PCR detection results showed as compared with groups B and C, E-cadherin protein and mRNA expression levels were significantly increased (P<0.01), c-Met and N-cadherin mRNA and protein levels were significantly decreased (P<0.01) in group A. There was no significant difference between groups B and C. (3) Scratch assay showed the scratch space was wide, and healing was inhibited in group A. the migration ability of MGC-803 cells in group A was reduced as compared with groups B and C (P<0.01), but there was no statistically significant difference in the scratch space between groups B and C. (4) Flow cytometry revealed that the apoptosis rate in groups A, B and C was (21.06±1.12)%, (2.18±0.14)% and (2.04±0.11%) respectively. The apoptosis rate in group A was significantly higher than in group C (P<0.01), and there was no significant difference between groups B and C. Conclusion The transfection of MACC-1 gene can promote the proliferation and inhibit the apoptosis of gastric cancer cell line MGC-803 cells, which may play a role in the regulation of hepatocyte growth factor (HGF)/c-Met signal transduction pathway. Key words: Gastric cancer; Colon cancer metastasis associated gene-1; Apoptosis; Biological function