AIM:To investigate the apoptosis-inducing effect ofCaspases-3 expressed by constructed eukaryotic vector ongastric cancer cell line SGC7901.METHODS:PCR was employed to amplify the sequencesof both small and large subunits of Caspases-3.Its productswere separately cloned into the Sma I site of pBluescriptKS+to generate both plasmids pBS/SS and pBS/LS.Thesmall subunit fragment was excised from plasmid pBS/SSwith BamH I and then inserted into the BamH I site of plasmidpBS/LS preceding that of the large subunit to yield plasmidpBS/Rev-Caspase-3.Rev-Caspase-3 cDNA was excised withKpn I+Xba I and then subcloned into plasmid pcDNA3.1(+)to construct Rev-Caspase-3 eukaryotic expression vectorpcDNA/Rev-Caspase-3,which was used to transientlytransfect SGC7901 cell line.Cell count,MTT assay and electronmicroscopy were used to confirm the antiproliferation andapoptosis-inducing effect of Rev-Caspase-3 expression ongastric cancer cells.RESULTS:Plasmid pBS/Rev-Caspase-3 and eukaryoticexpression vector pcDNA/Rev-Caspase-3 were successfullyconstructed.SGC7901 cells were transiently transfected byeither pcDNA/Rev-Caspase-3 or pcDNA3.1(+)for 24,48,72,and 96 h respectively.Cell growth was measured by cellcount and MTT assay.In cell count assay,the cell numberswere 1.8×10~6,1.55×10~6,2.0×10~6,and 3.1×10~6 in theexperimental group and 2.5×10~6,3.1×10~6,4.0×10~6,and5.7×10~6 in the control group at 24,48,72 and 96 h respectively.The growth of SGC7901 cells was suppressed by Rev-Caspase-3 in a time-dependent manner(P<0.05).The resultsof MTT assay were similar to that of cell count(P<0.05).The characteristics of apoptosis such as chromatincondensation,crescent formation and margination were seenand more obvious with time in the given-experimental periodin the experimental group,but not easily observed in thecontrol group.CONCLUSION:The expression of Rev-Caspase-3 by the constructed eukaryotic vector can significantly induceapoptosis of gastric cancer cell line SGC7901,which mayexhibit a potential way in gastric cancer gene therapy. AIM: To investigate the apoptosis-inducing effect of Caspases-3 expressed by constructed eukaryotic vector ongastric cancer cell line SGC7901.METHODS: PCR was employed to amplify the sequences of both small and large subunits of Caspases-3.Its products separately separately cloned into the Sma I site of pBluescript KS + to generate both plasmids pBS / SS and pBS / LS.Thesmall subunit fragment was excised from plasmid pBS / SSwith BamH I and then inserted into the BamH I site of plasmid pBS / LS preceding that of the large subunit to yield plasmid pBS / Rev-Caspase-3.Rev-Caspase-3 cDNA was excised with Kpn I + Xba I and then subcloned into plasmid pcDNA3.1 (+) to construct Rev-Caspase-3 eukaryotic expression vector pcDNA / Rev- Caspase-3, which was used to transientlytransfect SGC7901 cell line. Cell count, MTT assay and electronmicroscopy were used to confirm the antiproliferation andapoptosis-inducing effect of Rev-Caspase-3 expression ongastric cancer cells .RESULTS: Plasmid pBS / Rev- Caspase-3 and eukaryoticexpression vecto r pcDNA / Rev-Caspase-3 were successfully reconstructed. SGC7901 cells were transiently transfected by either pcDNA / Rev-Caspase-3 or pcDNA3.1 (+) for 24, 48, 72, and 96 h respectively. Cell growth was measured by cell count and MTT assay.In cell count assay, the cell numberswere 1.8 × 10 -6, 1.55 × 10 -6, 2.0 × 10 -6, and 3.1 × 10 -6 in the experimental group and 2.5 × 10 -6, 3.1 × 10 -6 , 4.0 × 10 -6, and 5.7 × 10 -6 in the control group at 24, 48, 72 and 96 h respectively. The growth of SGC7901 cells was suppressed by Rev-Caspase-3 in a time-dependent manner (P <0.05). The results of apoptosis such as chromatin condensation, crescent formation and margination were seen and more obvious with time in the given-experimental period in the experimental group, but not easily observed in the control group. CONCLUSION: The expression of Rev-Caspase-3 by the constructed eukaryotic vector can significantly induce apoptosis of gastric cancer cell line SGC7901, which may exhibi t a potential way in gastric cancer gene therapy.