摘要
目的探讨乙酰肝素酶反义硫代寡脱氧核苷酸对人肝癌细胞系侵袭力的抑制作用。方法设计合成互补于HPAmRNA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OligofectamineTMReagent包埋后转染SMMC-7721、BEL-7402细胞。采用RT-PCR和Western-blot检测转染前后细胞HPAmRNA及蛋白表达的变化,以基底膜重建实验检测SMMC-7721、BEL-7402细胞侵袭力的变化。结果与正常对照组相比,AS-ODN转染组SMMC-7721、BEL-7402细胞HPAmRNA和蛋白的表达均显著下降(P<0.01,P<0.05),转染后两细胞系的侵袭力抑制率分别为55.8%和61.8%。结论HPA硫代反义寡脱氧核苷酸可通过下调HPAmRNA和蛋白的表达抑制肝癌的侵袭力。
Objective To investigate the inhibitory effect of heparanase antisense oligodeoxynucleotide （AS-ODN） on the invasiveness of human liver cancer cell lines SMMC-7721, BEL-7402 in vitro. Methods We designed and synthesized AS-ODN that was complementary to the start codon region of heparanase mRNA, and the control, nonsense oligodeoxynucleotide （NS-ODN）. The ODNs with the final concentration of 300 nmol/L was delivered into SMMC-7721, BEL-7402 cells by Oligofectamine^TM Beagent. We evaluated heparanase gene expression using BT-PCR and detected heparanase protein expression using Western blot assay after transfection. Cell invasiveness was measured by Matrigel invasion assay. Results The result showed that heparanase gene expression, protein expression and invasiveness in AS-ODN group decreased significantly as compared with control groups （ P 〈 0. 05 ）. The inhibitory rate of AS-ODN was 55.8% in SMMC-7721 cell and 61.8% in BEL-7402 cell. Conclusion These resuits suggested that heparanase AS-ODN complementary to the start codon region significantly inhibit the invasiveness of human liver cancer cell lines SMMC-7721 and BEL-7402 by down regulating heparanase expression.
出处
《基础医学与临床》
CSCD
北大核心
2006年第7期734-738,共5页
Basic and Clinical Medicine
基金
河北省卫生厅科研基金(04029)
关键词
乙酰肝素酶
反义寡脱氧核苷酸
肝癌
侵袭力
heparanase
antisense oligodeoxynucleotide
hepatocarcinoma
invasiveness
