Construction and identification of psilencer3.0-HIF-? siRNA / 中国癌症杂志
China Oncology
; (12)2006.
Article
en Zh
| WPRIM
| ID: wpr-545722
Biblioteca responsable:
WPRO
ABSTRACT
Background and purpose:Carcinoma of the kidney is the most common malignant renal parenchymal carcinoma;its biological behavior is extremely complicated and is not sensitive to radiotherapy and chemotherapy.Studies have shown that the hypoxia-inducible factor HIF1-? and HIF2-? are related to the occurrence and development process of the clear-cell carcinoma of kidney.So,we intend to construct psilencer3.0-HIF-? siRNA recombinant plasmid in this study through RNA interference method,thereby providing an effective tool for further exploration of the role of HIF in the occurrence and development of clear-cell carcinoma of kidney.Methods:Design and chemically synthesize the DNA fragments of coding HIF-1? and HIF-2? siRNA and make them up into siRNA expression vector through gene recombination.Adopting the real-time quantitative PCR and Western blot tested the inhibitive effect of the constructed siRNA expression vector on mRNA and protein levels on the target gene expression.Results:After 786-0 cells transfecting with psilencer3.0-HIF-1?siRNA and psilencer3.0-HIF-2?,the inhibitory rate of mRNA expression of HIF-1? and HIF-2? reached 82.1% and 87.4% respectively,and OS-RC-2 cells transfecting psilencer3.0-HIF-1? siRNA and psilencer3.0-HIF-2?,the inhibition rate reached 91.2% and 81.2% respectively.The protein expression of the experimental group with 786-0 cells and OS-RC-2 cells transfecting HIF-1? interference plasmid and HIF-2? interference plasmid were lower at different levels than that from the blank control group.Conclusions:The constructed siRNA expression vector can effectively inhibit the expression of target gene HIF-1 and HIF-2 at mRNA and protein levels.
Texto completo:
1
Índice:
WPRIM
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Idioma:
Zh
Revista:
China Oncology
Año:
2006
Tipo del documento:
Article