Construction of Recombinant Caspase-3 Gene and the Observation of It's Apoptotic Activity in Pancreatic Carcinoma

Objective To explore the new gene therapy method for tumor, the recombinant Caspase 3 gene (r caspase 3) eukaryotic expression plasmid was constructed by molecular biologic method. Methods The eukaryotic expression plasmid pcDNA3.1(+)/r Caspase 3 was constructed by rearrangement of the large subunit and small subunit of Caspase 3 and it was transfected into pancreatic carcinoma cells(PC Ⅱ). After being transfected, the expression of r Caspase 3 mRNA in pancreatic carcinoma cells was detected by RT PCR and it's apoptotic activity was detected by FCM. Results The sequence of r Caspase 3 showed that the recombinant molecules (r Caspase 3) now had its' small subunit preceding its' large subunit. After pancreatic carcinoma cells being transfected with the pcDNA3.1(+)/r Caspase 3 by liposomes, a 894 bp strap was observed by RT PCR. No strap was found in control groups. A transparent hypodiploid karyotype peak was found by FCM.Conclusion The plasmid of pcDNA3.1(+)/r Caspase 3 has been constructed successfully. r Caspase 3 has apoptotic activity and can be used as target gene in gene therapy for pancreatic carcinoma.