Title: Changes of MMP-2 and TIMP-2 Expressions on Cultured Human Pancreatic Cancer Cell Line PCNA-1 Transfected with Smad4 cDNA
Abstract: Objective The purpose of the present study was to elucidate the mechanism of the role of Smad4 in the invading and metastasis of pancreatic cancer by observing matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2(TIMP-2) expressions on cultured human pancreatic cancer cell line PCNA-1 transfected with wild-type Smad4 cDNA.Methods Lipofectin method was used to transfect wild-type Smad4 cDNA into PCNA-1 and transfected cells were selected with G418,reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blot analysis for detecting Smad4 mRNA and protein expression level.The expressions of MMP-2 and TIMP-2 were also determined by RT-PCR and Western blot.Results Stable overexpression of Smad4 on two transfected PCNA-1 clones(F-9,F-21) were successfully established.Stable transfection of wild-type Smad4 cDNA into the PCNA-1 cell line resulted in decreased expressions of MMP-2 mRNA and its protein(about 62% lower),accompanied with increased synthesis of TIMP-2 mRNA and its protein(about 2.1-fold) compared with the control group.Conclusion These results suggest that Smad4 gene transfection may inhibit spreading and invasion of pancreatic cancers cells,which would play an important role in the anti-metastasis therapy for pancreatic cancer.
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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