乙酰肝素酶反义寡核苷酸对裸鼠人肺癌细胞皮下移植瘤的抑制作用  

Inhibitory effects of heparanase antisense oligodeoxynucleotide on human lung cancer cells transplanted subcutaneously in nude mice

作　　者：陈志涛[1] 祝慧珍[2] 朱良明[1] 方玉松[1] 田辉[3] 

机构地区：[1]山东大学附属济南市中心医院胸外科,山东济南250013 [2]山东大学附属济南市中心医院胃肠外科,山东济南250013 [3]山东大学齐鲁医院胸外科,山东济南250012

出　　处：《中国现代医学杂志》2012年第29期6-10,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:30571844);山东省自然科学基金(No:2009ZRB14005)

摘　　要：目的探讨乙酰肝素酶反义寡核苷酸(HPSE ASODN)对人肺癌A549细胞裸鼠皮下移植瘤的抑制作用。方法设计合成互补于HPSE mRNA起始密码区的HPSE ASODN,以阳离子脂质体Lipofectamine包埋后转染人肺癌A549细胞进行培养。实验分ASODN组、脂质体组和对照组,采用Western-blotting检测各组A549细胞HPSE蛋白的表达;将反义寡核苷酸(ASODN)转染成功的人肺癌A549细胞注射于裸鼠皮下复制移植瘤模型(ASODN组),绘制移植瘤生长曲线,以免疫组织化学方法检测移植瘤肿瘤微血管密度(MVD),比较3组间肿瘤体积和MVD的差异。结果与对照组和脂质体组比较,ASODN组A549细胞HPSE蛋白的表达受到明显抑制,差异有统计学意义(P<0.01);ASODN组裸鼠成瘤时间晚,移植瘤体积明显小于脂质体组和对照组,差异有统计学意义(P<0.01)。ASODN组、脂质体组和对照组移植瘤的MVD计数分别为(13.5±1.8)、(24.3±2.5)和(24.7±2.6),与脂质体组、对照组比较,ASODN组MVD计数明显降低,差异有统计学意义(P<0.01)。结论 HPSE ASODN明显抑制裸鼠人肺癌移植瘤的生长和血管生成,有希望成为一种有效的肺癌基因治疗方法。【Objective】 To study the inhibitory effects of heparanase antisense oligonucleotide（HPSE ASODN） on the growth and angiogenesis of tumor xenografts derived from A549 human lung cancer cell lines in nude mice.【Methods】 HPSE ASODN which was complementary with initiation codon region of HPSE mRNA was designed and synthesized.After embedded by cation lipofectamine,it was transfected into A549 cells of human lung cancer.The inhibitory efficient of HPSE ASODN was analyzed by examination of HPSE protein expression using western blotting.The cells were separated into three groups as follows： ASODN group（transfected with the lipofectamine and ASODN）,lipofectamine group（transfected with lipofectamine only） and control group.Cells were injected subcutaneously into nude mice to establish the model of human lung cancer xenografts respectively.The growth curve of the tumor xenografts was observed.Furthermore,microvessel density（MVD） of the tumor xenografts in each group was detected by immunohistochemistry.【Results】 Western blot showed the expression of HPSE protein in ASODN group was remarkably down-regulated compared with the control group and the lipofectamine group（P〈0.01）.The formation of tumor xenografts in the ASODN group was apparently late,and the volume of the tumor was smaller than that in the control group and lipofectamine group（P〈0.01）,the inhibitory rate was 64.3% at the time of the sixth week.The MVD counts were（13.5±1.8）,（24.3±2.5）,（24.7±2.6） in ASODN,lipofectamine and control group respectively.Compared to control（24.7±2.6） and lipofectamine group（24.3±2.5）,the MVD counts in ASODN group（13.5±1.8） was apparently decreased（P〈0.01）.【Conclusion】 The heparanase antisense oligonucleotide can inhibit the tumor growth and angiogenesis of the xenografts of human lung cancer cell lines A549 in nude mice.

关 键 词：肺肿瘤 乙酰肝素酶 反义寡核苷酸 微血管密度 

分 类 号：R734.2[医药卫生—肿瘤]