[Construction of recombinant caspases-3 gene and the test of its apoptotic activity in pancreatic carcinoma cell strain].

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.

• Authors: Wei Wang, Lixin Liu, Zhiguo Liu, Lunan Yan
• Journal: Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi (Sheng Wu Yi Xue Gong Cheng Xue Za Zhi) Vol. 20 Issue 4 Pg. 671-4 (Dec 2003) ISSN: 1001-5515 [Print] China
• PMID: 14716874 (Publication Type: English Abstract, Journal Article)

Chemical References

• CASP3 protein, human
• Caspase 3
• Caspases

Topics

• Apoptosis
• Caspase 3
• Caspases (biosynthesis, genetics, physiology)
• Cloning, Molecular
• Genetic Therapy
• Humans
• Pancreatic Neoplasms (pathology, therapy)
• Plasmids (genetics)
• Transfection
• Tumor Cells, Cultured