Effect Of RASSF1A Gene On EMT And Nude Mice Xenograft Tumor In Human Gastric Cancer Cell Line

Objective:Based on the preliminarystudies of research group, we build Nude mice transplantation tumor model of, human gastric cancer cell lines to further study the effect of RASSF1 A gene on EMT. Methods:1) Western Blottechnology observe the expression of EMT related proteins like E-cadherin、Vimentin and Snail protein that in human gastric carcinoma cell line MGC-803 transfected RASSF1 A gene or not.2)Building Nude mice transplantation tumor model of human gastric cancer cell lines SGC-7901 and MGC-803, each nude mice inoculatedhuman gastric cancer cell lines are divided into three part: thecells without transfection、the cells transfected with pcDNA3.1(+)、the cells transfected with pcDNA3.1(+)-RASSF1 A plasmid. Observe the growing of transplantation tumor, drawing the transplantation tumor’ growth chart diagram, Stripping xenograft tumor after 4-6 weeks, Compares the transplantation tumor’ weight and volume, than HE staining.Using Western Blottechnology observe the expression of EMT related proteins: E-cadherin、Vimentin and Snail protein, witch come from xenograft tumor proteins. Results:1) By stabily transfected RASSF1 A gene, that human gastric cancer MGC-803 cells ’western blot results showed this: the protein of epithelial marker E – cadherin had high expression 、the protein of vimentin、snail had lower expression in RASSF1A/ pcDNA3.1(+)/MGC-803 cell group, compared with MGC-803 cell group、pcDNA3.1(+)/MGC-803 cell group, compare among groups had difference in statistic sence(P < 0.05). While between MGC-803 cell group and pc DNA3.1(+)/MGC-803 cell group, it had no obvious difference(P > 0.05).2) The results showed that the nude mice xenograft tumor model with stably high expression of RASSF1 A gene in human gastric cancer SGC- 7901 cells and MGC- 803 cells was successfully established, nude mice xenograft tumor’volume、weight of the grop transplanted positive plasmid were significantly less than the grop transplanted empty plasmid 、 not transplanted group, the difference was statistically significant(P < 0.05). While betweenthe grop transplanted empty plasmid、not transplanted group, it had no obvious difference(P > 0.05); Under the microscope lens it showed, nude mice xenograft tumor of the grop transplanted positive plasmid had less atypia than the grop transplanted empty plasmid、not transplanted group. Western blot results showedthat the protein of epithelial marker E-cadherin had high expression、the protein of vimentin、snail had lower expression in the grop transplanted positive plasmid,compared with the grop transplanted empty plasmid、not transplanted group, compare among groups had difference in statistic sence(P < 0.05). While between the grop transplanted empty plasmid、not transplanted group, it had no obvious difference(P > 0.05). Conclusion:1)RASSF1A gene can significantly inhibit the ability of gastric cancer cell turning into a tumor.2)RASSF1A gene can inhibit the occurrence of EMT in human gastric cancer SGC- 7901 and MGC-803 cells.