Nonspecific apoptosis induced by long, double-stranded RNA in human osteosarcoma cells
- VernacularTitle:长双链RNA非特异性诱导人骨肉瘤细胞凋亡的实验研究
- Author:
Zhen WANG
;
Min YANG
;
Liwen LI
- Publication Type:Journal Article
- Keywords:
Osteosarcoma;
Cell death, programmed;
Gene expression regulation
- From:
Chinese Journal of Orthopaedics
1999;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the plasmid that can produce long,double-stranded RNA and identify nonspecific apoptosis induced by long, double-stranded RNA in human osteosarcoma cells. Methods Plasmid pCI-neo-dsRNA was constructed by two promoters driven DNA sequence based on nontoxic, completely exoteric gene EGFP from the head and the end simultaneously to synthesize sense and antisense RNAs. Human osteosarcoma cells MG63 were transfected with pCI-neo-dsRNA and 48 h later the transfected cells were assessed by Apoptosis Detection System, FCMand MTT method. Results Plasmid pCI-neo-dsRNA was constructed successfully, MTT measurement showed that the absorbance of cells decreased gradually with the increase of plasmid pCI-neo-dsRNA dose(0.05 ?g, 0.10 ?g, 0.15 ?g, 0.20 ?g,0.25 ?g,0.30 ?g). MG3 transfected with pCI-neo-dsEGFP presented a great amount of irregular green fluorescence that indicated cell apoptosis. As the quantity of dsRNA increased, the percent of apoptosis cells also raised. While the dose of pCI-neo-dsEGF was 3.0 ?g, the apoptotic results of pCI-neo-dsEGF in MG3 were similar to that of 5-FU. Conclusion Long, double- stranded RNA could trigger human osteosarcoma cell apoptosis and the further research on using long, double-stranded RNA in osteosarcoma gene therapy can be done based on this theory.
