摘要
目的 观察谷胱甘肽 S 转移酶P1(GSTP1)启动子调控的胞嘧啶脱氨酶和尿嘧啶磷酸核糖转移酶融合基因 (CD UPP ,即自杀基因 )表达的腺病毒载体 ,对卵巢癌顺铂耐药细胞的体外杀伤作用。方法 采用细菌内同源重组法构建腺病毒载体 ,设立卵巢癌顺铂敏感细胞株A2 780和以其诱导的耐药细胞株A2 780 /DDP ,在体外用重组腺病毒转染两组细胞并联合应用前药 5 氟胞嘧啶 (5 FC) ,观察两组卵巢癌细胞对 5 FC的敏感性。将CD UPP阳性和CK UPP阴性的A2 780 /DDP细胞按不同比例混合后给予 5 FC ,观察 5 FC对混合细胞的杀伤作用。结果 当重组腺病毒滴度为 10 0感染复数(MOI)、5 FC浓度为 2 5 0 μg/ml时 ,对顺铂耐药的A2 780 /DDP细胞的存活率为 (3 6± 1 0 ) % ,对顺铂敏感的A2 780细胞的存活率为 (76 5± 2 8) % ,两者比较 ,差异有显著性 (P <0 0 5 )。此外 ,2 0 %CD UPP阳性A2 780 /DDP细胞 ,即可引起 80 3%的混合细胞死亡 ,CD UPP / 5 FC系统表现出明显的旁观者效应。结论 由腺病毒介导的含有GSTP1调控的CD UPP/ 5 FC系统 ,对卵巢癌顺铂耐药细胞具有特异性杀伤作用 ,为临床上克服卵巢癌化疗耐药 ,提供了一条安全、高效的治疗途径。
Objective To construct an adenoviral vector in which the fusion gene cytosine and uracil phosphoribosyl (UPP) transferase was directed by glutathione S-transferase P1(GSTP1 ) promoter, and to investigate specific killing effect of the suicide gene system on cisplatin-resistant ovarian cancer cells. Methods Recombinant adenovirus was generated through homologous recombination in bacteria. A2780 and A2780/DDP cells were infected with Ad and then received flucytosine (5-FC) administration. The relative survival of these cells was tested. And a bystander effect was observed by mixing gene-transferred and gene-untransferred A2780/DDP cells with 5-FC. Results In vitro, when MOI was 100 and 5-FC was 250 μg/ml, relative survival rate of A2780/DDP cells was only (3.6±1.0)%;that of A2780 cells was (76.5±2.8)%. Significant bystander effect was caused by CD-UPP gene and 20% gene-transferred A2780/DDP cells induced 80.3 % of total cells to death. Conclusion Recombinant adenovirus carrying CD-UPP gene driven by GSTP1 promoter has a specific killing effect on cisplatin-resistant ovarian cancer cells.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2004年第7期478-481,i004,共5页
Chinese Journal of Obstetrics and Gynecology
基金
国家自然科学基金资助项目 (3 0 0 70 786)