Effect Of RASSF1A Gene On Migration And Invasion In Human Gastric Cancer SGC-7901Cells

Objective: The aim of this study is to investigate the influence of RASSF1A geneon migration and invasion and its molecular mechanism in human gastric carcinomacell line SGC-7901.Methods: The transient transfection method is used to transfect human gastriccancer SGC-7901cells, for verifying the transfection effect，RT-PCR and WesternBlot are used to detect the expression of RASSF1A gene after transfecting48hours.Through cell scratch experiment and Transwell invasion experiment discussmigration and invasion in human gastric carcinoma cell line SGC-7901transfectedRASSF1A gene or not, respectively. For further study, Western Blot is employed toobserved the expression of E-cadherin, Vimentin and Snail protein in human gastriccarcinoma cell line SGC-7901transfected RASSF1A gene or not.Results:RT-PCR and Western Blot showed that the mRNA and protein of RASSF1Agene had high expression in SGC-7901cells transfected with pcDNA3.1(+)-RASSF1A plasmid, compared with SGC-7901cells without transfection andSGC-7901cells transfected with pcDNA3.1(+), and the difference was statisticallysignificant(P <0.05).While between the controls, the SGC-7901cells withouttransfection and SGC-7901cells transfected with pcDNA3.1(+) had no obviousdifference (P>0.05). It was prompted that RASSF1A gene was successfullytransfected to human gastric cancer SGC-7901cells. Then, Scratch experimentsdemonstrated that the mobility of SGC-7901cells transfected with pcDNA3.1(+)-RASSF1A plasmid was obviously lower, compared with SGC-7901cellstransfected with pcDNA3.1(+) and SGC-7901cells without transfection, and thedifference was statistically significant (P <0.05). While between SGC-7901cells transfected with pcDNA3.1(+) and SGC-7901cells without transfection, there wereno significant difference(P>0.05). It was indicated that RASSF1A gene could inhibitthe migration ability in human gastric cancer SGC-7901cells. Transwell experimentproved that the membrane cells was significantly reduced in SGC-7901cellstransfected with pcDNA3.1(+)-RASSF1A plasmid, compared with the controlgroups(P <0.05), prompted that the invasion ability was abate. There were nosignificant changes between SGC-7901cells transfected with pcDNA3.1(+) andSGC-7901cells without transfection(P>0.05). It was suggested that RASSF1A genecould inhibit the invasion ability in human gastric cancer SGC-7901cells. In order toclarify the molecular mechanisms, we further studied on the protein of epithelialmarkers E-cadherin, mesenchymal markers Vimentin and transcription factor Snail,found that E-cadherin was up-regulated in pcDNA3.1(+)-RASSF1A/SGC7901group,while Vimentin and Snail protein were decline, compared with the blank controlgroup and pcDNA3.1(+)/SGC7901group(P <0.05). Whereas, between the untreatedgroup and pcDNA3.1(+) plasmid group, no statistically significant difference (P>0.05), prompted that RASSF1A gene can inhibit the occurrence of EMT in humangastric cancer SGC-7901cells.Conclusion:RASSF1A gene can inhibit migration and invasion in human gastric cancerSGC-7901cells, its mechanism may be related to RASSF1A gene down-regulated theexpression of transcription factor Snail, thereby inhibiting epithelial mesenchymalrelated transformation in human gastric cancer SGC-7901cells.