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Inhibitory effect of siRNA on HPA gene expression in SKOV3 cells
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Abstract:
Objective: To study the inhibitory effect of siRNA on Heparanase (HPA) expression in SKOV3 cells. Methods:Two pairs of 21 bp reverse repeated sequence targeting HPA RNA (spaced by 9 bp nucleotide) were synthesized and were cloned into plasmid Pgenesil-1 to construct recombinant plasmid PGenesil-1(+)-HPA expressing 2 hairpin siRNAs. The inhibition of HPA gene was detected by RT-PCR, real-time PCR and immunohistochemical staining after PGenesil-1(+)-HPA was stably transfected into SKOV3 cells. Results: The recombinant plasmid PGenesil-1(+)-HPA (expressing 2 hairpin siRNAs) was successfully constructed. RT-PCR, real-time PCR and immunohistochemical staining showed that there was a significant decrease in HPA mRNA and protein level in experimental group compared with those in control group. Conclusion: siRNA targeting HPA mRNA can specically suppress the expression of HPA gene in SKOV3 cells; RNA interference method provides a new way for studying the role of HPA and gene therapy of cancer.
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