Construction of recombinant caspases-3 gene and the test of its apoptotic activity in pancreatic carcinoma cell strain.
- Author:
Wei WANG
1
;
Lixin LIU
;
Zhiguo LIU
;
Lunan YAN
Author Information
1. Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Caspase 3;
Caspases;
biosynthesis;
genetics;
physiology;
Cloning, Molecular;
Genetic Therapy;
Humans;
Pancreatic Neoplasms;
pathology;
therapy;
Plasmids;
genetics;
Transfection;
Tumor Cells, Cultured
- From:
Journal of Biomedical Engineering
2003;20(4):671-674
- CountryChina
- Language:Chinese
-
Abstract:
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.
