重组腺病毒载体介导的人野生型p53基因增强肝癌细胞对化疗药物的敏感性  

Enhanced Chemosensitivity to Anticancer Agents in Hepatocellular Carcinoma Cell Lines Transduced with Recombinant Adenoviral Vector Expressing Wild-Type Human p53 Genes in vitro

作　　者：施明[1] 王福生[1] 刘明旭[1] 张冰[1] 雷周云[1] 邱兆华[2] 高兰兴[3] 吴祖泽[2] 

机构地区：[1]解放军302医院生物工程研究室,北京100039 [2]军事医学科学院放射医学研究所 [3]军事医学科学院卫生学环境医学研究所

出　　处：《中国肿瘤生物治疗杂志》2001年第2期80-83,共4页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金 ( 39970 831)资助

摘　　要：目的 :观察表达人野生型p5 3 ,GM CSF和B7 1基因的重组腺病毒载体 (BB 10 2 )提高肝癌细胞对化疗药物的敏感性。方法 :BEL 74 0 2 ,HLE和HuH 73株肝癌细胞被 5 0MOI的BB 10 2转染后 ,Westernblot检测p5 3基因的表达 ,MTT方法检测肝癌细胞对化疗药物的敏感性。结果 :当MOI为 5 0pfu/细胞时 ,腺病毒载体对 3株肝癌细胞的转染效率均达到 80 %以上。转染BB 10 2后 ,HLE和HuH 7细胞对顺铂的敏感性分别提高 11倍和 2 8倍 ,对丝裂霉素 C的敏感性分别提高 3 .75倍和 2 0倍。而BB 10 2的转染对BEL 74 0 2细胞的化疗敏感性没有影响。结论 :BB 10 2腺病毒载体转染后能显著提高p5 3基因突变的HLE和HuH 7细胞对顺铂和丝裂霉素 C的敏感性 ,因而增强顺铂和丝裂霉素 C对HLE和HuHObjective: To investigate the reversal of multidrug resistance (MDR) in hepatocellular carcinoma cell lines mediated by transfer of adenoviral vector (designed as BB 102) expressing human wild type p53,GM CSF and B7 1 in in vitro cell system. Methods: Three human hepatocellular carcinoma cell (HCC) lines: BEL 7402 with wild type p53, HLE and HuH 7 with mutant p53, were transduced with BB 102 at the 50 pfu/cell of MOI dose. The expression level of human wild type p53 gene was determined by Western blot analysis. The sensitivity of HCC cells to anticancer agents was evaluated by MTT assay. Results: The human wild type p53 was highly expressed in all the BB 102 transduced HCC cell lines at a MOI of 50 pfu/cell. The MTT assay showed that the ID 50 for cisplatin decreased from 2.2 μg/ml to 0.2 μg/ml for HLE cells and from 28 μg/ml to 1 μg/ml for HuH 7 cells, respectively. The ID 50 for mitomycin C simultaneously decreased from 1.2 μg/ml to 0.32 μg/ml for HLE cells and from 0.1 μg/ml to 0.005 μg/ml for HuH 7 cells, which reversed the chemosensitivity of resistant HLE and HuH 7 cell lines. There was no significant change for ID 50 in the BB 102 transduced BEL 7402 cells.Conclusion: Recombinant adenoviral vector BB 102 could efficiently transduce into HCC cells and express human genes including p53, which restores the chemosensitivity of p53 mutant tumor cells to anticancer agents.

关 键 词：重组腺病毒 P53基因 肝细胞癌 顺铂 丝裂霉素-C 药敏试验 

分 类 号：R735.7[医药卫生—肿瘤] R730.53[医药卫生—临床医学]