六味地黄丸对自杀基因系统杀伤肝癌细胞增效作用的量效关系  

Dose-Effect Study on Enhancing Effect of Liuwei Dihuang Bolus on the Killing Effect of Suicide Gene Therapy for Hepatoma Cells

作　　者：杜标炎[1] 张爱娟[2] 谭宇蕙[3] 易华[1] 吴映雅[3] 郭玉荣[2] 

机构地区：[1]广州中医药大学病理教研室,广东广州510405 [2]广州中医药大学 [3]广州中医药大学生物化学教研室,广东广州510405

出　　处：《广州中医药大学学报》2009年第1期59-62,共4页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:30572433)

摘　　要：【目的】观测六味地黄丸含药血清对自杀基因HSV-tk/GCV系统杀伤大鼠肝癌CBRH7919细胞的增效作用,探讨建立自杀基因联合中药方药的肿瘤基因治疗联合方案的可行性。【方法】构建HSV-tk/GCV自杀基因治疗系统,并确定丙氧鸟苷(GCV)的工作浓度(39.2μmol/L);制备SD大鼠4 d和8 d六味地黄丸(剂量为32.gkg-.1d-1)灌胃含药血清和生理盐水灌胃空白血清;用大鼠肝癌细胞CBRH7919/tk-或tk+与tk-1∶9比例混合细胞按3×103个/孔铺96孔板,分为空白血清组、GCV加空白血清组、10%tk+/GCV加空白血清组、含药血清组、GCV加含药血清组及10%tk+/GCV联合含药血清组;含药血清又再分为低剂量组、中剂量组和高剂量组,每组6个复孔,用完全培养基培养24 h后,各组分别加入相应空白血清或含药血清,孵育12 h,加入终浓度39.2μmol/L的GCV,培养60 h,采用四甲基偶氮唑盐(MTT)法检测细胞活性,Q值法分析自杀基因系统与含药血清联合的相互作用是否具有协同性。【结果】空白血清组、含药血清组、GCV加空白血清组、GCV加含药血清组细胞均未显示明显的细胞毒性。10%tk+/GCV加空白血清组及10%tk+/GCV联合不同浓度的含药血清组均具有显著杀伤作用(与空白血清组比较,P<0.05),联合组存活率显著低于10%tk+/GCV加空白血清组(P<0.05);含药血清中剂量和高剂量组的联合作用具有协同性(Q>1.15),且有浓度依赖性。给药4 d血清和给药8 d血清结果无显著性差异(P>0.05)。【结论】六味地黄丸含药血清对自杀基因系统10%tk+/GCV杀伤大鼠肝癌CBRH7919细胞具有协同增效作用,且有一定的量效关系。Objective To investigate the possibility of establishing a combined regimen of Chinese medicine and suicide gene therapy,we observed the enhancing effect of medicated serum of Liuwei Dihuang Bolus（LDB） on the killing effect of HSV-tk/GCV suicide gene system for rats hepatoma cell line CBRH7919.Methods The HSV-tk/GCV suicide gene therapy system was constructed and the working solution of ganciclovir（GCV） at 39.2μmol/L was detected firstly.Then medicated serum was prepared from SD rats treated with LDB（32g·kg-1·d-1）by gavage for 4 and 8 days,respectively,and the non-medicated serum（blank serum） was prepared from SD rats treated with normal saline.CBRH7919/tk＋ cells or CBRH7919/tk-cells were mixed with tk-in proportion of 1∶9 and then the mixture was used as 10%tk＋/GCV target cells.CBRH7919/tk-cells or 10%tk＋ cells were seeded in quadruplicate in 96-well plates at a density of 3×103 /well,grown in complete medium for 24 hours,treated with medicated or non-medicated serum for 12 hours and sequentially with GCV at 39.2μmol/L for another 60 hours.In terms of the added serum and GCV treatment,the CBRH7919 cells were sorted into following groups：non-medicated serum group（10% non-medicated serum,final concentration,V∶V）,GCV plus non-medicated serum group,non-medicated serum plus 10%tk＋/GCV group,low-dose medicated serum group（2.5% medicated serum＋ 7.5% non-medicated serum）,GCV plus low-dose medicated serum group,low-dose medicated serum＋10%tk＋/GCV group,mid-dose medicated serum group（5% medicated serum＋ 5% non-medicated serum）,GCV plus mid-dose medicated serum group,mid-dose medicated serum plus 10%tk＋/GCV group,high-dose medicated serum group（10% medicated serum）,GCV plus high-dose medicated serum group,high-dose medicated serum plus 10%tk＋/GCV group.Six bias-wells were set for each group.The viability of CBRH7919 cells was determined by MTT assay.Q value analysis was used to evaluate the synergistic effect of the medicated serum on the suicide gene system.Th

关 键 词：六味地黄丸/药理学 肝肿瘤/中西医结合疗法 基因治疗 肿瘤细胞 培养 

分 类 号：R285.5[医药卫生—中药学]