Experimental study on relationship between Smad4 and angiogenesis related factors in human gastric cancer cell line / 中华消化杂志

ABSTRACT

Objective To study the effect of Smad4 gene on angiogenesis related factors in human gastric cancer cell line.Methods Recombinant eukaryotie expressing plasmid pcDNA3.1 (-)-Smad4 containing Smad4 gene and empty vector pcDNA3.1 (-) were introduced into human gastric cancer cell line MKN28 using lipofectam and selected by G418,respectively.Two cell lines were obtained as followsSmad4+-MKN28 cell line which was MKN28 transfected with a stable hybrid containing Smad4 gene and Smad4--MKN28 cell line with empty plasmid as control.The transcription and expression of VEGF and TSP1 were investigated by RT-PCR and Western blot.Results The mRNA expression of TSP1 in Smad4+-MKN28 cells was higher (P＜0.05) than that in control cells,while VEGF was lower(P＜0.05).Western blot showed the consistent results as measurement by RT-PCR.Conclusion Smad4 restoration in gastric cancer cells reduced angiogenesis rates through down-regulation of angiogenesis activitor and up-regulation of angiogenesis inhibitor.