乙酰肝素酶反义硫代寡核苷酸对人肝癌细胞珠BEL-7402侵袭力的影响  

Inhibitory effect of heparanase antisense oligodeoxynucleotides on invasiveness ofBEL-7402 cell line

作　　者：王顺祥[1] 吴晓慧[1] 周少英[1] 彭利[1] 

机构地区：[1]河北医科大学第四医院肝胆外科,河北石家庄050011

出　　处：《中华肿瘤防治杂志》2006年第5期344-346,350,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨乙酰肝素酶(Hpa)反义硫代寡脱氧核苷酸(AS-ODN)对人肝癌细胞BEL-7402侵袭力的影响。方法:设计合成互补于HpamRNA起始密码区的AS-ODN,以阳离子脂质体Oligofectami-neTMReagent包埋后转染BEL-7402细胞。采用RT-PCR和Western-blot检测不同浓度AS-ODN转染后细胞HpamRNA及蛋白表达的变化,以基底膜重建实验检测细胞侵袭力的变化。结果:0、100、200和300nmol/LAS-ODN转染组BEL-7402细胞HpamRNA的表达量分别为0·911、0·898、0·774和0·280;Hpa65×103蛋白表达量分别为0·923、0·875、0·834和0·237;Hpa50×103蛋白表达量分别为1·872、1·919、0·821和0·325。300nmol/LAS-ODN转染后BEL-7402细胞HpamRNA和蛋白的表达明显下降;0、100、200和300nmol/LAS-ODN转染组侵袭细胞数分别为137±15、141±13、134±18和49±7,300nmol/L,AS-ODN转染组侵袭细胞数较正常对照组明显减少,P=0·000。结论:一定浓度的HpaAS-ODN可通过下调HpamR-NA和蛋白的表达抑制肝癌细胞的侵袭力。OBJECTIVE,To investigate the inhibitory effect of heparanase antisense oligodeoxynucleotide （AS-ODN） on the invasiveness of human liver cancer cell line BEL-7402 in vitro. METHODS, AS-ODN complementary to the start codon region of heparanase mRNA respectively was designed and synthesized. The ODNs with the final concentration of 0 nmol/L, 100 nmol/L, 200 nmol/L, 300 nmol/L were delivered into BEL-7402 cells by Oligofectamine^TM Reagent. After transfeeticn the heparanase gene and protein expression level were evaluated by the reverse transcription polymerase chain reaction （RT-PCR） and Western blot respectively and the cell invasiveness was measured by the Matrigel invasion assay. RESULTS： The heparanase gene expression in 0, 100, 200, 300 nmol/L AS-ODN transfeeted groups were 0. 911, 0. 898, 0. 774, 0. 280, respeetively; 65 × 10^3 Hpa protein expressions were 0. 923, 0.875, 0. 834, 0. 237 respeetively; 50 × 10^3 Hpa protein expressions were 1.872, 1.919, 0. 821, 0. 325, respectively. The heparanase gene expression, protein expression in 300 nmol/L AS-ODN transfeeted group decreased compared with the control group. Invasion cells in 0, 100, 200, 300 nmol/L AS-ODN transfeeted groups were 137 ± 15, 141±13, 134 ± 18, 49 ± 7, respectively. Invasion cells in 300 nmol/L AS-ODN transfeeted group decreased significantly compared with the normal control group,P=0. 000. CONCLUSIONS： Heparanase AS-ODN complementary to the start codon region could significantly inhibit the invasiveness of human liver cancer cell line BEL-7402 by down regulating heparanase expression.

关 键 词：肝肿瘤/病理学 葡糖醛酸糖苷酶 寡棱苷酸类 反义 肿瘤侵润 

分 类 号：R735.7[医药卫生—肿瘤]