低氧时反义寡核苷酸对胰腺癌乙酰肝素酶表达及侵袭力的影响  

Effects of antisense oligodeoxynucleotide on expression of heparanase mRNA and invasion of pancreatic carcinoma in hypoxia

作　　者：孟珂伟[1] 吴武军[2] 宋占文[1] 周先亭[1] 王雪峰[1] 张永明[1] 李绍军[1] 

机构地区：[1]青岛大学医学院附属烟台毓璜顶医院肝胆外科,烟台264000 [2]陕西省人民医院肝胆外科,西安710061

出　　处：《中华内分泌外科杂志》2010年第3期148-151,共4页Chinese Journal of Endocrine Surgery

基　　金：山东省烟台市科学技术发展计划项目（2006131.15）

摘　　要：目的 研究低氧对胰腺癌乙酰肝素酶（Hpa）表达的影响及与肿瘤细胞侵袭力的关系.方法 体外低氧培养细胞,观察低氧在不同时间胰腺导管癌细胞株BxPC-3细胞Hpa mRNA和蛋白表达的变化;以Tranwell侵袭小室实验观察低氧及应用反义寡核苷酸（AS-ODN）阻断Hpa表达后肿瘤细胞侵袭力的变化,明确低氧促进肿瘤细胞侵袭力增加是否与Hpa有关.结果 在常氧条件下,BxPC-3细胞Hpa mRNA和蛋白即有低量表达.在低氧条件下,Hpa mRNA和蛋白表达在3 h受到轻度抑制,在6 h、12 h、24 h和48 h表达明显升高,且mRNA和蛋白的表达水平一致.预先以Hpa AS-ODN序列阻断Hpa的表达,则低氧不能诱导Hpa mRNA和蛋白表达上调.低氧使穿过Transwell小室的BxPC-3细胞数量增加了96.2%（P〈0.01）,Hpa AS-ODN（400 nmol/L）对低氧诱导的侵袭细胞的抑制率为37.2%,差异有统计学意义（P〈0.05）.结论 低氧可通过上调胰腺癌BxPC-3细胞Hpa mRNA和蛋白的表达,提高肿瘤细胞侵袭力.Objective To study the regulation of heparanase expression by hypoxia and its correlation to the invasiveness of tumor cells. Methods BxPC-3 cells were cultured in hypoxia in vitro and the heparanase mRNA and protein expression were detected by reverse transcriptional polymerase reaction chains （RT-PCR） and western blot respectively. Matrigel invasion assay was used to observe the invasive abilities of tumor cells in hypoxia and in the status of heparanase was inhibited by antisense oligodeoxynucleotide （AS-ODN） targeting to the heparanase gene promoter. Results In normoxia, there was a relatively low level of heparanase mRNA and protein expression in cultured BxPC-3 cells. In hypoxia, heparanase expression, mRNA and protein which expressed consistently, were inhibited slightly at 3h and upregulated significantly at 6h, 12h, 24h and 48h. When the heparanase expression was inhibited by AS-ODN, the heparanase mRNA and protein maintained low in hypoxia, however, the nonsense oligodeoxynucleotide （NS-ODN） did not block upregulation of heparanase expression. In matrigel assay, after 48h incubation, number of BxPC-3 cells that penetrated the Matrigel-coated filter of transwell chamber was increased 96.2% in hypoxia （P〈0.01）, the Hpa AS-ODN （400 nmol/L） inhibited the invasive cells by 37.2% （P〈0.05）. Conclusions BxPC-3 cells invasion ability is enhanced by hypoxia through upregulation of heparanase mRNA and protein expression in BxPC-3 pancreatic cancer cell lines.

关 键 词：胰腺癌 乙酰肝素酶 反义寡核苷酸类 低氧 

分 类 号：R735.9[医药卫生—肿瘤]