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[Lentivirus-mediated gene transfer of human sTRAIL induced apoptosis in SGC-7901 cells].
- Source :
-
Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition [Sichuan Da Xue Xue Bao Yi Xue Ban] 2013 May; Vol. 44 (3), pp. 348-51, 392. - Publication Year :
- 2013
-
Abstract
- Objective: To construct self-inactivating (SIN) lentiviral vector carrying human soluble tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL) gene and to observe the effects of sTRAIL gene on apoptosis in SGC-7901 cells, so as to assess the value of sTRAIL in gene therapy for gastric cancer.<br />Methods: The bicistronic SIN lentiviral transfer plasmid containing sTRAIL gene and internal ribosomal entry site-green fluorescent protein gene (IRES-GFP) was constructed. Recombinant lentivirus containing sTRAIL were packaged using liposome by lentiviral packing system. Several cell lines including HL-7702, HLF-1 and SGC-7901 cells were infected with the viral supernatant. Flow cytometry was used to measure apoptosis of cells and recombinant sTRAIL. protein was assayed by ELISA at 24 h after infection.<br />Results: The lentiviral transfer plasmid pXZ208-sTRAIL was constructed, and the virus titres were above 10(6) IU/mL in the supernatant. SGC-7901 cells were efficiently infected by recombinant virus. The apoptosis rate of SGC-7901 cells was increased with the virus multiplicity of infection (MOI) increasing. When the MOI was > or = 0.5, a dose-dependent apoptosis rate was observed. At MOI of 6.0, the highest apoptosis rate (29.12 +/- 2.87)% was observed, while the expression of sTRAIL in the supernatant was only (34.08 +/- 3.43) ng/mL. However, no significant apoptosis was observed in HL-7702 cells and HLF-1 cells.<br />Conclusion: Recombinant lentivirus carrying human sTRAIL gene can efficiently infected SGC-7901 cells, which induced apoptosis in SGC-7901 cells in vitro by secreting bioactive sTRAIL protein. However, this effect is not seen in normal cells.
- Subjects :
- Cell Line, Tumor
Gene Transfer Techniques
Genetic Vectors genetics
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
Humans
Lentivirus metabolism
Stomach Neoplasms metabolism
TNF-Related Apoptosis-Inducing Ligand biosynthesis
Apoptosis genetics
Lentivirus genetics
Stomach Neoplasms pathology
TNF-Related Apoptosis-Inducing Ligand genetics
Transfection
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1672-173X
- Volume :
- 44
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition
- Publication Type :
- Academic Journal
- Accession number :
- 23898510