Effects of shRNA targeting maspin on invasion of gastric carcinoma SGC7901 cell line.
Maspin is a member of the serpin (serine protease inhibitor) family of protease inhibitors known to have tumor suppressor activity in diverse human cancers. However, maspin gene function and the molecular aspects in gastric carcinoma remain largely unclear. To investigate the effects of maspin on invasion of gastric carcinoma SGC7901 cell line and the underlying molecular mechanism involved in this process, we cloned short hairpin oligoes (shRNA) targeting maspin into plasmid pGenesil-1.1 eukaryotic expression vector and then transfected the recombinant plasmid pGenesil-maspin into gastric carcinoma SGC7901 cells using Lipofectamine 2000. After the maspin expression was successfully knocked down, the number of cells invading through Matrigel was obviously increased (P<0.05) in the Transwell chamber assay. By detection of reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis, respectively, we found that mRNA and protein of uPA, VEGF-C were increased significantly, and the protein level of MMP7 was also increased (P<0.05). These results suggested that maspin gene could inhibit invasion of gastric cancinoma SGC7901 cells and this inhibition maybe result from the interaction between maspin and uPA, MMP7, or VEGF-C.
Department of Pathology, Molecular Medicine and Tumor Center, Chongqing Medical University, Chongqing, P.R. China.
Yaying
Y
Yang
Haipeng
H
Shi
Xiujun
X
Li
Yongfen
Y
Yi
D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D055785:Gene Knockdown Techniques; D006801:Humans; D020783:Matrix Metalloproteinase 7 / Q000378:metabolism; D009361:Neoplasm Invasiveness; D034741:RNA, Small Interfering; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015843:Serpins / Q000235:genetics / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D014162:Transfection; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism
D034741:RNA, Small Interfering; C085344:SERPIN-B5; D015843:Serpins; D042582:Vascular Endothelial Growth Factor C; D014568:Urokinase-Type Plasminogen Activator; D020783:Matrix Metalloproteinase 7