TK/GCV系统对骨肉瘤MG-63细胞杀伤的体外实验研究  

Experimental research in vitro of TK/GCV system for osteosarcoma MG-63 cell damage

作　　者：张桦栋[1] 吕智[1] 冯毅[1] 刘小丽[1] 侯慧铭[1] 

机构地区：[1]山西医科大学第二医院骨科,山西太原030001

出　　处：《中国骨伤》2014年第3期240-243,共4页China Journal of Orthopaedics and Traumatology

基　　金：山西省自然科学基金资助项目(编号:2007011112)~~

摘　　要：目的:探讨脂质体介导的TK/GCV系统对骨肉瘤MG-63细胞的杀伤作用以及所产生的旁观者效应。方法:脂质体介导TK基因体外转染骨肉瘤MG-63细胞,倒置荧光显微镜观察细胞转染是否成功,流式细胞仪检测转染细胞与未转染细胞的转染效率。将未转染的骨肉瘤MG-63细胞分为3组,实验1组用转染TK/GCV的细胞上清液与原培养液按1/10、1/7、1/5、1/2比例混和液培养;实验2组用0.22μm滤器过滤的转染后的细胞上清液与原培养液按1/10、1/7、1/5、1/2比例混和液培养;对照组用培养液培养,四甲基偶氮唑盐(MTT)比色法分别测定各组细胞的生长抑制率及骨肉瘤细胞对TK/GCV系统的敏感性。结果:经TK基因转染的MG-63细胞,倒置荧光显微镜下有大量绿色荧光蛋白表达,流式细胞仪检测转染TK基因的细胞转染效率可达75.5%。6 d后MTT检测结果显示实验1组中各比例浓度的混合培养液对细胞的抑制率与对照组相比有统计学意义(P<0.05);实验2组中1/10、1/7比例浓度的混合培养液对细胞的抑制率与对照组相比差异无统计学意义(P>0.05)。经TK基因转染的MG-63细胞随GCV浓度增加,细胞凋亡率增加。结论:脂质体介导的TK/GCV系统能够通过旁观者效应抑制骨肉瘤MG-63细胞的生长。Objective：To study the killing effects of the liposome mediated thymidine kinase （TK）/ganciclovir （GCV） system on MG-63 osteosarcoma （OS） cells and its bystander effects. Methods：Liposome mediated TK gene transfected into MG-63 OS cells,the efficiency of transfection was analyzed by flow cytometry and observed under inverted fluorescence mi-croscope. Non-transfected osteosarcoma MG-63 cells were divided into three groups ,in the experimental group 1 transfected TK/GCV cells cultured in solutiona liquid mixture by supernatant by 1/10 ,1/7,1/5,1/2 ratio to original broth;in the experi-mental group 2 transfected cells cultured in solutiona liquid mixture of supernatant filtered through 0.22 μm filter by 1/10,1/7,1/5,1/2 ratio to original broth,in control group the transfection cells cultured in original culture solution. Cell growth inhi-bition rate and osteosarcoma cell sensitivity to TK / GCV system were measured by MTT assay in each group. Results：The TK gene was transfected into MG-63 OS cells successfully by liposome mediated,flow cytometry instrument detection TK gene transfection cell transfection efficiency can reach 75.5%. Six days later the MTT assay showed that in the experimental group 1 inhibition rate of all concentration ratio of the mixed culture fluid were statistically significant as compared with the control group （P〈0.05）,and in the experimental group 2 that of the 1/10 and 1/7 of concentration ratio of mixed culture medium was not statistically significant as compared with the control group （P〉0.05）. TK gene transfected MG-63 cells increased with the the GCV concentration,the cell apoptosis rate increased. Conclusion：The experiment demonstrated that the MG-63 OS cells are sensitive to the liposome mediated TK/GCV system and bystander effects are significant.

关 键 词：骨肉瘤 基因治疗 胸腺嘧啶脱氧核苷激酶(TK) 丙氧鸟苷(GCV) 旁观者效应 

分 类 号：R687[医药卫生—骨科学]