The Expression And Function Study Of EMMPRIN In SGC-7901

Objective: To construct the recombinant plasmids ofpEGFP-N1-EMMPRIN and pshRNA-EMMPRIN, transfected into humangastric cancer SGC-7901cells by lipofectamine2000, to examine theirexpression in eukaryotic cell SGC-7901and to detect its role on invasionand migration of human gastric cancer SGC-7901cells, to ay thefoundation for further study on the gene therapy of tumor with EMMPRIN.Methods:(1) To extract total mRNA from human colon cancer, thehuman EMMPRIN gene was obtained by RT-PCR and cloned intopEGFP-N1vector, and then the recombinant plasmid ofpEGFP-N1-EMMPRIN was transfected into human gastric cancerSGC-7901cells by lipofectamine2000, and the expression in SGC-7901cells were detected by fluorescent microscopy, RT-PCR and Western blot;(2)To construct pshRNA-EMMPRIN against human EMMPRIN. Theinterference plasmid of the EMMPRIN was screened by sequencing,RT-PCR and western blot;(3) Transfected into human gastric cancerSGC-7901cells by lipofectamine2000with pEGFP-N1-EMMPRIN andpshRNA-EMMPRIN. Transewell assay tested the effect of EMMPRIN on invasion and migration of human gastric cancer SGC-7901cells.Results:(1) The EMMPRIN cDNAs were obtained by RT-PCR, therecombinant plamids pEGFP-N1-EMMPRIN were constructed successfully.Transfected it into SGC-7901cells, it could be expressed and the fusionprotein in SGC-7901cells were observed;(2) The pshRNA-EMMPRINwere constructed and the most interference plasmid of the EMMPRIN wasscreened by sequencing, RT-PCR and western blot.(3) Significantdifference on invasion and migration of human gastric cancer SGC-7901cells was observed between the group pEGFP-N1-EMMPRIN and thegroup pshRNA-EMMPRIN.Conclusion: EMMPRIN can increase the ability which invasion andmigration of tumor cells. On the contrary, reduce EMMPRIN expressioncan significantly relieve the invasion and migration of tumor cells, lay thefoundation for further study on the gene therapy of tumor with EMMPRIN.