Screening of effective sequence of heparanase gene-targeted by small interfering RNA / 中国现代普通外科进展

ABSTRACT

Objective: According to heparanase’s gene sequence of gene bank, to construct heparanase gene-targeted small interfering RNA (siRNA)and its expression vector and to observe its interfering effect on the expression of heparanase gene in human malignant breast cancer MDA-MB-231 cells. Methods: Heparanase gene-targeted hairpin siRNA was designed, two complementary oligonucleotide strand was synthesized and inserted into pGPU6/GFP/Neo vector,which was identified by sequence identify. Human malignant breast cancer MDA-MB-231 cells were transfected with the constructed vector with lipofectamine method. Western blot was per-formed to evaluate the expression of heparanase protein. Results: Four kinds of heparanase gene-targeted hairpin siRNA were designed, and were inserted into pGPU6/GFP/Neo vector after annealing. The vector containing siRNA was proved to be right by sequencing. The result of Western blot indicated that the expression of heparanase could be degraded by siRNA. Conclusion: The expression of heparanase can be degraded by siRNA method, and HPSE-A and HPSE-B showed the best results.