摘要
目的 探讨 5 -氟胞嘧啶 ( 5 FC )热化疗对转染组织特异性胞嘧啶脱氨基酶 (CD )基因的大肠癌细胞SW 480的作用。方法 采用脂质体法将CEA基因顺式转录调控序列 (TRS )驱动CD基因的组织特异性逆转录病毒载体G 1CEACDNa转导入大肠癌细胞SW 480 ,以G 418筛选阳性克隆扩增后 ,采用水浴加温法 ,43℃作用 3 0min共 3次 ,同时给予 5 FC进行敏感试验 ;RT PCR法检测目的基因在靶细胞的表达。结果 SW 480 CEACD细胞在 43℃作用 3 0min共 3次条件下 ,CD基因能稳定表达 ;热疗本身对SW 480细胞有一定的杀伤作用 ( P <0 .0 5 ) ,转CD基因后 ,SW 480细胞对 5 FC的敏感性明显提高 (P <0 .0 1) ,热疗与前药 5 FC合用 ,对SW 480 CEACD的杀伤作用显著大于对未转基因细胞的杀伤作用 ( P <0 .0 1) ,亦大于单独应用 5 FC时对SW 480 CEACD细胞的杀伤作用(P <0 .0 5 ) ,热化疗增加了SW 480 CEACD细胞对 5 FC的敏感性 ,并可观察到明显的旁观者效应。结论 热疗与 5 FC联合应用 ,提高了CEA组织特异性CD /5 FC系统对结肠癌细胞SW 480的靶向性杀伤作用。
Objective To investigate the effect of thermochemotherapy with 5 fluorocytosine(5 FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue specific cytosine deaminase (CD) in vitro.Methods Recombinant retroviral vector G1CEACDNa, in which CD gene was controlled under the CEA promoter, was introduced through liposome technique to the human colorectal carcinoma cell line SW480, and then the cells were selectively cultured in G418. The proliferated colonies were treated with the combined therapy of 5 FC and hyperthermia at a 43℃ over 30 min for 3 times. RT PCR was performed to analyze the expression of CD gene in target cells after thermochemotherapy. Results The CD gene expressed steadily in the target cells after 3 times of hyperthermic treatment. After the transfection of CD gene, SW480 CEACD cells were more sensitive to 5 FC than their parental cells (P < 0.01), and the killing effect of hyperthermia on SW480 cells could also observed (P < 0.05). Furthermore , after treated with thermochemotherapy of 5 FC at 43℃ over 30 min, the killing effect on SW480 CEACD cells was more obvious than that on SW480 cells(P < 0.01). 5 FC combining with hyperthermia showed a higher anti tumor effect on SW480 CEACD cells than 5 FC alone did(P< 0.05 ). The onlooker effect in the cells transfected with CD gene was obviously observed. Conclusions Hyperthermia can improve the anti tumor effect of 5 FC on human colorectal carcinoma cell lines SW480 transfected with the CEA tissue specific CD genes.
出处
《中国普通外科杂志》
CAS
CSCD
2003年第6期427-430,共4页
China Journal of General Surgery
