[Construction of heparanase gene-targeted small interfering RNA and its gene silencing effect].
Zhejiang Da Xue Xue Bao Yi Xue Ban
; 36(6): 581-7, 2007 11.
Article
en Zh
| MEDLINE
| ID: mdl-18067232
OBJECTIVE: To construct heparanase gene-targeted small interfering RNA(siRNA) and its expression vector and to observe its interference effect on the expression of heparanase gene in human malignant melanoma A375 cell. METHODS: Heparanase gene-targeted hairpin siRNA was designed, two complementary oligonucleotide strand was synthesized and inserted into pRNATU6.1 vector, which was then identified by PCR and sequencing. Human malignant melanoma A375 cells were transfected with the constructed vector using lipofectamine method. Semi-quantitative PCR was performed to evaluate the heparanase-mRNA expression levels, and Western blot was performed to evaluate the expression of heparanase protein. RESULTS: The vector containing siRNA was identified by PCR and sequencing; the results of semi-quantitative PCR and Western blot showed that the expression levels of both heparanase RNA and protein in transfected A375 cells were decreased significantly(P<0.05). CONCLUSION: The heparanase gene-targeted siRNA and its vector were successfully constructed, which can reduce the heparanase gene and protein expression in transfected cells.
Buscar en Google
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Regulación Neoplásica de la Expresión Génica
/
ARN Interferente Pequeño
/
Interferencia de ARN
/
Glucuronidasa
/
Melanoma
Límite:
Humans
Idioma:
Zh
Revista:
Zhejiang Da Xue Xue Bao Yi Xue Ban
Asunto de la revista:
MEDICINA
Año:
2007
Tipo del documento:
Article
País de afiliación:
China
Pais de publicación:
China