Inhibitory effect of siRNA on HPA gene expression in SKOV3 cells / 中国肿瘤生物治疗杂志

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 nueleotide) were synthe- sized and were cloned into plasmid PGenesil-1 to construct recombinant plasmid PGenesil-1(+)-HPA expressing 2 hair- pin 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.