ABSTRACT
Objective To investigate the inhibitory effect of maspin gene on metastasis and invasion of gastric cancer and its mechanism.Methods Maspin gene was ligated to the expression vector PCR2.1 with T4 DNA ligase after the PCR2.1 was digested by HindⅢ/XbaⅠ.The recombinant vector maspin/PCR2.1 was transfected into human gastric cancer cell lines MKN28 and SGC7901,then the mRNA and protein expression changes of maspin gene,uPA and uPAR were detected respectively by PT-PCR and Western blotting.Results After identified by digestion and sequencing,the reconstructed plasmid was confirmed to contain the correct and full nucleotide sequence of maspin gene.The expressions of maspin gene were up-regulated in MKN28 and SGC7901 cell lines,while the expressions of uPA and uPAR were down-regulated.Conclusion The expression vector maspin/PCR2.1 is constructed successfully and can be expressed in eukaryotic cells.Expressions of uPA and uPAR can be inhibited by maspin gene.
ABSTRACT
Objective:To construct the expression vector maspin /Pcr2.1 in order to provide the evidence for the inhibitory effect of maspin gene on growth,metastasis and invasion of hepatocellular carcinoma(HCC) and to discuss its mechnism.Methods:The full length of maspin gene was cloned,and the expression vector PCR2.1 was digested by Hind111/XbaI,then the maspin gene and the PCR2.1 vector were ligated with T4 DNA ligase to be recombinant vector.The recombinant vector maspin/PCR2.1 was transfected into humman hcc MHCC-97 cell line,and the expressive changes of maspin gene were detected.Results:The recombinant plasmid was amplified in the E.coli.JM109.After the identification of redigesting by HindIII/XbaI and sequencing,the reconstructive plasmid was confirmed to contain the correct and full nucleotide sequence of maspin gene,and the mRNA and protein level of maspin gene were up-regulated in hcc MHCC-97 cell.Conclusion:The expression vector maspin/PCR2.1 is constructed successfully and can be expressed in eukaryotic cells.