Zhang Hua-Dong, Lu Zhi, Feng Yi, Liu Xiao-Li, Hou Hui-Ming
Zhongguo Gu Shang. 2014 Mar;27(3):240-3.
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.
Liposome-mediated TK gene transfected into MG-63 OS cells, the efficiency of transfection was analyzed by flow cytometry and observed under inverted fluorescence microscope. 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 experimental group 2 transfected cells cultured in solutiona liquid mixture of supernatant filtered through 0.22 microm 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 inhibition rate and osteosarcoma cell sensitivity to TK/GCV system were measured by MTT assay in each group.
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.
The experiment demonstrated that the MG-63 OS cells are sensitive to the liposome-mediated TK/GCV system and bystander effects are significant.
研究脂质体介导的胸苷激酶（TK）/更昔洛韦（GCV）系统对MG-63骨肉瘤（OS）细胞的杀伤作用及其旁观者效应。
将脂质体介导的TK基因转染至MG-63骨肉瘤细胞，通过流式细胞术分析转染效率，并在倒置荧光显微镜下观察。未转染的骨肉瘤MG-63细胞分为三组，实验组1中，将转染TK/GCV的细胞培养上清液与原培养液按1/10、1/7、1/5、1/2比例混合培养；实验组2中，将转染细胞培养上清液经0.22μm滤膜过滤后与原培养液按1/10、1/7、1/5、1/2比例混合培养，对照组则在原培养液中培养。采用MTT法检测各组细胞生长抑制率及骨肉瘤细胞对TK/GCV系统的敏感性。
脂质体成功介导TK基因转染至MG-63骨肉瘤细胞，流式细胞仪检测TK基因转染细胞的转染效率可达75.5%。六天后MTT法检测显示，实验组1各浓度比例混合培养液的抑制率与对照组相比差异均有统计学意义（P<0.05），而实验组2中浓度比例为1/10和1/7的混合培养液与对照组相比差异无统计学意义（P>0.05）。TK基因转染的MG-63细胞随着GCV浓度增加，细胞凋亡率升高。
实验表明MG-63骨肉瘤细胞对脂质体介导的TK/GCV系统敏感，且旁观者效应显著。
