Experimental study on the Caspase-3 siRNA decreases apoptosis in stable cell lines of bone marrow mesenchymal stem cells
10.3969/j.issn.1674-7445.2014.05.005
- VernacularTitle:Caspase-3 siRNA减少骨髓间充质干细胞稳定株细胞凋亡的实验研究
- Author:
Libao LIU
1
;
Lei HUANG
;
Junhang ZHANG
;
Hongying LIAO
;
Ailing HU
;
Shilei XU
;
Ping HUA
Author Information
1. 中山大学附属第三医院心胸外科
- Keywords:
Caspase-3;
RNAinterference;
Retrovirus;
Cellapoptosis;
Bonemarrowmesenchymal stem cell
- From:
Organ Transplantation
2014;(5):283-288
- CountryChina
- Language:Chinese
-
Abstract:
Objective ToexploreamethodofusingCaspase-3smallinterferenceribonucleicacid (siRNA)in vitro to decrease the apoptosis in stable cell lines of bone marrow mesenchymal stem cell (MSC)in ordertoestablishMSCcelllineswithstableexpression.Methods VirusparticlescontainingCaspase-3siRNA were generated in 293FT cells by retroviral packaging system,then were transfected into MSC of rats. And the transfected cells were screened and cultured to get the stable MSC lines. According to the characteristics of retrovirus,the stable expression of the cell lines was identified by Western blot and real-time fluorescent quantitative PCR (RT-QPCR). The apoptosis of stable cell lines and normal MSC were detected by terminal dexynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL ) and their differences were compared.Results Comparedwiththenormalcells,thecaspase-3proteinexpressionandmRNAcontentof stable MSC lines were significantly reduced. In the same condition in vitro,the apoptosis quantity of stable MSC linessignificantlydecreasedcomparedwiththenormalcells.Conclusions StablecelllinesofMSCwithstable expression of Caspase-3 siRNA can be obtained by retroviral packaging system. The apoptosis quantity of stable MSC Lines significantly decreased compared with the normal cells.
