[Construction of recombinant caspases-3 gene and the test of its apoptotic activity in pancreatic carcinoma cell strain].
To explore the new gene therapeutic method for pancreatic carcinoma, the recombinant Caspases-3 gene (r-Caspases-3) was constructed by molecular biologic method. The eukaryotic expression plasmid pcDNA 3.1 (+)/r-Caspase-3 was constructed by rearrangement of the large subunit and small subunit of caspases-3, and then it was transfected into pancreatic carcinoma cells strain (PC-II). After being transfected, the expression of r-Caspase-3 mRNA in pancreatic carcinoma cells was detected by RT-PCR and its apoptotic activity was detected by FCM. The sequencing of the recombinant molecules (r-Caspases-3) confirmed that its small subunit preceded its large subunit. After the pancreatic carcinoma cells were transfected with the pcDNA3.1(+)/r-Caspases-3 by liposomes, an 894 bp strap was observed by means of RT-PCR. No strap was found in control groups. A transparent hypodiploid karyotype peak was revealed by FCM. The above data indicate that the gene of 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.
Wang
Wei
W
Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041.
Liu
Lixin
L
Liu
Zhiguo
Z
Yan
Lunan
L
D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D015316:Genetic Therapy; D006801:Humans; D010190:Pancreatic Neoplasms / Q000473:pathology* / Q000628:therapy; D010957:Plasmids / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases