乙酰肝素酶反义寡核苷酸对人肝癌细胞株乙酰肝素酶表达的影响

The effect of heparanase antisense oligodeoxynucleotides on heparanase gene expression of human liver cancer cell line

目的:探讨乙酰肝素酶(Hpa)反义寡脱氧核苷酸对人肝癌细胞株Hpa mRNA和蛋白表达的影响。方法:设计合成互补于Hpa mRNA起始密码区的硫代反义寡脱氧核苷酸AS-ODN及相应的无义对照寡脱氧核苷酸NS-ODN,以阳离子脂质体Oligofectamine^(TM) Reagent包埋后转染SMMC-7721、BEL-7402细胞,以RT-PCR和Western-blot检测转染前后细胞Hpa mRNA以及蛋白表达的变化。结果:与正常对照组和NS-ODN相应浓度组相比,转染AS-ODN组肝癌细胞Hp amRNA和蛋白的表达均下降。结论:Hpa反义寡脱氧核苷酸下调Hpa mRNA和蛋白的表达,可能抑制肝癌的侵袭转移。

Objective To investigate the effects of heparanase antisense oligodeoxynucleotide(AS-ODN) on the heparanase gene expression of human liver cancer cell lines(SMMC-7721 and BEL-7402) in vitro. Methods AS-ODN complementary to the start codon region of heparanase mRNA and its controlled nonsense oligodeoxynucleotide(NS-ODN) was designed and synthesized. The ODNs with the final concentration of 300 nmol/L were delivered into SMMC-7721 and BEL-7402 cells by OligofectamineTM reagent. After transfection,the heparanase gene and protein expression levels were evaluated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot,respectively. Results The heparanase gene and protein expression levels in AS-ODN group decreased significantly as compared with those in control groups. In contrast,those in NS-ODN group were not significantly different from those in liposome control group and in normal control group. Conclusion Heparanase AS-ODN complementary to the start codon region could significantly inhibit the invasiveness of human liver cancer cell lines(SMMC-7721 and BEL-7402) by down-regulating heparanase expression.