ODC和AdoMetDC双反义腺病毒载体抑制人食管癌Eca109细胞增殖和侵袭的研究  

Ornithine Decarboxylase and S-adenosylmethionine Decarboxylase Bi-antisense Virus Inhibit Growth and Invasion of Esophageal Cancer Cell Line Eca109

作　　者：宋旭[1] 田辉[1] 刘贤锡[2] 张冰[2] 李文军[1] 徐杰[1] 

机构地区：[1]山东大学齐鲁医院胸外科,山东济南250012 [2]山东大学医学院分子生物学实验中心,山东济南250012

出　　处：《癌症》2008年第11期1144-1149,共6页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.30571844);山东大学博士后基金项目(2005)~~

摘　　要：背景与目的:食管癌是我国常见的恶性肿瘤之一,对人民的生命和健康危害极大。有研究发现,在食管癌细胞和肿瘤组织中多胺的含量和生物合成明显增高。本研究中我们探讨ODC和AdoMetDC双反义腺病毒载体(Ad-ODC-AdoMetDCas)对人食管癌Eca109细胞多胺合成、细胞增殖和侵袭的抑制作用。方法:采用活细胞计数法和BrdU掺入实验观察Ad-ODC-AdoMetDCas对Eca109细胞增殖的影响;应用Westernblot法和HPLC法分别检测腺病毒对Eca109细胞中ODC和AdoMetDC蛋白表达的影响,以及胞内多胺合成的抑制作用,采用Matrigel侵袭实验分析腺病毒载体对Eca109细胞侵袭活性的影响,同时检测Ad-ODC-AdoMetDCas对Eca109细胞裸鼠皮下移植瘤生长的抑制作用。结果:活细胞计数法和BrdU掺入实验表明,Ad-ODC-AdoMetDCas对Eca109细胞增殖有明显的抑制作用(P<0.05)。Westernblot证实Ad-ODC-AdoMetDCas可明显抑制Eca109细胞中ODC和AdoMetDC基因表达。HPLC结果显示,感染Ad-ODC-AdoMetDCas后,Eca109细胞内三种多胺腐胺、精脒、精胺含量均明显降低(P<0.05)。Matrigel侵袭实验结果显示,Ad-ODC-AdoMetDCas可显著降低Eca109细胞的体外侵袭能力(P<0.05)。同时发现,与Ad-GFP相比,Ad-ODC-AdoMetDCas对裸鼠皮下移植瘤具有明显的抑制作用。结论:ODC和AdoMetDC双反义腺病毒载体具有显著抑制Eca109细胞增殖和侵袭的作用。BACKGROUND ＆ OBJECTIVE： Esophageal carcinoma is one of the most common malignant tumors in China. It is reported that the content and biosynthesis of polyamine in esophageal cancer is markedly increased. This study was to investigate inhibitory effects of both antisense ornithine decarboxylase （ODCas） and S-adenosylmethionine bi-antisense （AdoMetDCas） on polyamine biosynthesis, cell proliferation and invasion of esophageal cancer cell line Eca109. METHODS： An adenoviral vector Ad-ODC- AdoMetDCas containing antisense sequences of ODCse and AdoMetDCas was used. Cell proliferation was observed by counting viable cells or BrdU labeling. Protein expressions of ODC and AdoMetDC and the polyamine content in Eca109 cells were measured by Western blot and high performance liquid chromatography （HPLC）, respectively. Invasion of Ecal09 cells in vitro was detected using Matriget invasion assay. Furthermore, the anti-proliferation effect of Ad-ODC-AdoMetDCas on Eca109 cell xenografts in nude mice was evaluated. RESULTS： Ad-ODC-AdoMetDCas significantly inhibited proliferation of Eca109 cells （P〈0.05） and protein expressions of ODC and AdoMetDC. Transfection of Ad-ODC-AdoMetDCas significantly decreased synthesis of three polyamines in Eca109 cells, which were putrescine （Put）, spermidine （Spd） and spermine （Som） （P〈0.05）. Ad-ODC-AdoMetDCas dramatically suppressed invasiveness of Eca109 cells in vitro （P〈0.05）. In addition, compared with Ad-GFP, Ad-ODC-AdoMetDCas significantly suppressed the growth of Eca109 cell xenografts in nude mice. CONCLUSION.. Ad-ODC-AdoMetDCas significantly inhibits cell proliferation and invasion in esophageal cancer Eca109 cells.

关 键 词：鸟氨酸脱羧酶 S-甲硫氨酸脱羧酶 多胺 ECA109 细胞 增殖 侵袭 抑制作用 

分 类 号：R735.1[医药卫生—肿瘤]