野甘草醇对胸腺激酶依赖的丙氧鸟苷阻断膀胱癌进展的增效作用  

The synergistic effect of scopadulciol in treating human bladdr cancer by thymidine kinase-dependent ganciclovir

作　　者：瞿长宝[1] 蔡文清[1] 王圆圆[2] 王亚轩[1] 王东彬[1] 杨书文[1] 黎玮[1] 齐进春[1] 

机构地区：[1]河北医科大学第二医院泌尿外科,石家庄050000 [2]河北医科大学第二医院内科教研室,石家庄050000

出　　处：《中华实验外科杂志》2013年第9期1912-1914,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察野甘草醇（SDC）在人端粒酶逆转录酶（hTERT）启动子调控腺病毒介导的单纯疱疹病毒胸腺激酶基因/丙氧鸟苷（HSV-TK/GCV）自杀基因系统对人膀胱癌细胞的体外杀伤中的增效作用.方法 测定胸苷激酶（TK）活性,应用高效液相色谱（HPLC）法测定三磷酸丙氧鸟苷（GCV-TP）在肿瘤细胞中的浓度,按转染倍数（MOI）=100加入Ad-hTERT-HSV-TK,24 h后分别加入5％血清培养液含GCV（200、100、50、10、5、l、0 mg/L）,含SDC（0、0.04或0.20 μmol/L）,应用于膀胱癌253J细胞,观察对膀胱癌细胞的杀伤作用.旁杀伤效应观察：将Ad-hTERT-HSV-TK按MOI=100转染253J细胞,24 h后将转染后的253J细胞和未转染的253J细胞以不同比例混合培养,共分11组（1∶1、1∶2、1∶3、1∶4、2∶1、2∶3、3∶1、3∶2、3∶4、4∶1、4∶3）,在11组253J肿瘤细胞培养液中分别加入GCV使其浓度为100μmoL/L或浓度为100 μmol/L的GCV联合应用SDC 0.04μmoL/L,噻唑蓝（MTT）测定细胞存活率,观察旁杀伤作用.结果 SDC对转染Ad-hTERT-HSV-TK的膀胱癌253J细胞的TK酶活性影响不明显.应用高效液相色谱法分析,以单加GCV组的GCV-TP浓度记作100％.加入0.04 μmol/L SDC后,GCV-TP浓度变化差异无统计学意义（P＞0.05）；加入5倍治疗剂量0.20 μmol/L SDC后,GCV-TP浓度提高到（105.0±5.0）％,差异有统计学意义（P＜0.05）.进一步细胞实验证实,SDC对Ad-hTERT-HSV-TK/GCV系统杀伤膀胱癌253J细胞具有增效作用,旁杀伤作用增强明显.结论 SDC对胸腺激酶依赖的GCV阻断膀胱癌进展有一定增效作用,将天然药物SDC与构建的具有肿瘤特异性的重组腺病毒Ad-hTERT-HSV-TK/GCV系统联合应用,可以达到减毒增效的目的.Objective To investigate the enhanced effect of dulciol （SDC） on the killing effect on antitumor actions by Ad-human telomerase reverse transcriptase （hTERT）-herpes simplex virus thymidine kinase （HSV-TK）/ganciclovir（GCV） suicide gene system on bladder cancer cells in vitro.Methods To clarigy the mechanism of enhanced effect of SDC in the gene therapy of human bladder cancer cell line 253J by thymidine kinase （TK）-denendent GCV in molecular level,the TK enzyme activity and the concentration of GCV-TP in the tumor cells were measured by using high performance liquid chromatography （HPLC）.The bladder cancer cells,respectively,were treated with different doses and different concentrations Ad-hTERT-HSV-TK [multiplicity of infection （MOI） =100],GCV （200,100,50,10,5,1,0 mg/L） and SDC （0,0.04 or 0.20 μmol/L） with The killing effect.The bystander effect of these drugs in therapy of human bladder cancer cells were observed by treated with different mixing proportion of TK ＋ cells andTK-cells （1∶1,1∶2,1∶3,1∶4,2∶1,2∶ 3,3∶1,3∶2,3∶4,4∶1,4∶3）.Results SDC had slightly effect on the TK enzyme activity of bladder cancer cells that were transduced by Ad-hTERT-HSV-TK.The GCV-TP concentration in the GCV treatment group was 100％.There was no significant difference when 0.04 μmoL/L SDC was added,The GCV-TP concentration was up to （105.0 ± 5.0） ％ when adding 0.20 μmol/L SDC,and there was significant difference campared to the GCV treatment group （P ＜0.05）.It was confirmed that SDC had strong killing effect on the bladder cancer cells by combination of Ad-hTERT-HSV-TK and GCV,the bystander effect was also significantly augmented by the combined treatment of GCV and SDC.Conclusion The synergistic effect of SDC is significantly shown in the therapy of human bladder cancer cells by TK-dependent GCV.SDC is effective in the Ad-hTERT-HSV-TK/GCV administration system in killing bladder cancer cells.

关 键 词：膀胱肿瘤 基因治疗 端粒酶逆转录酶 重组腺病毒 

分 类 号：R737.14[医药卫生—肿瘤]