SLC22A18基因转染人胶质瘤U251细胞对放疗敏感性的影响  

Influence of introducing SLC22A18 gene on the radiosensitivity of human glioma U251 cells

作　　者：楚胜华[1] 马延斌[1] 冯东福[1] 邱建华[1] 张红[1] 朱志安[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院神经外科,上海201900

出　　处：《现代肿瘤医学》2012年第3期466-469,共4页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:30901535);上海交通大学医学院"新百人计划"资助项目(编号:10XBR01);上海交通大学医学院附属新华医院集团科研资助项目(编号:10XHJT01)

摘　　要：目的:探讨SLC22A18基因转染人胶质瘤U251细胞对放疗敏感性的影响。方法:用脂质体介导的转染技术将pIRES2-EGFP-SLC22A18表达重组质粒导入U251细胞,通过逆转录-聚合酶链反应(RT-PCR)和免疫印迹(Western blot)检测SLC22A18基因和蛋白的表达。U251细胞分为4组:对照组、转染组、放疗组和转染联合放疗组。集落形成实验检测各组U251细胞集落形成数,用四甲基偶氮唑盐微量酶反应比色法(MTT法)和流式细胞仪检测各组U251细胞生长抑制率和凋亡率,进一步应用裸鼠皮下荷瘤模型观察脂质体介导的pIRES2-SLC22A18表达重组质粒转染U251细胞对放疗敏感性的影响。结果:重组质粒转染组通过RT-PCR和Western blot证实了SLC22A18基因和蛋白在U251细胞中表达。集落形成实验检测发现SLC22A18基因对U251细胞的集落形成数为(60.6±5.2)个,当放射剂量为3、6和9 Gy时,U251细胞的集落形成数分别为(30.0±3.6)、(13.0±3.0)和(4.0±1.0)个。MTT检测发现SLC22A18基因对U251细胞的生长抑制率为(80.12±5.75)%。当放射剂量为3、6和9 Gy时,U251细胞的生长抑制率分别为(17.05±4.24)%、(17.34±1.62)%和(18.71±4.59)%。当SLC22A18基因与放疗(3、6和9 Gy)联合作用时,抑制率分别为(81.45±5.32)%、(90.45±1.65)%和(92.62±2.12)%。SLC22A18基因转染所产生的U251细胞凋亡率为17.68%。放疗(3、6和9 Gy)引起的细胞凋亡率分别为4.64%、4.87%和5.42%。当SLC22A18基因与放疗(3、6和9 Gy)联合作用时,凋亡率分别为18.42%、21.48%和23.92%。体内实验结果显示,SLC22A18基因对U251细胞6 Gy照射的抑瘤率比较为1.81。结论:SLC22A18基因转染增加了人胶质瘤U251细胞对放疗的敏感性。Objective：To study the effect of SLC22A18 gene on the radiosensitivity of human glioma U251 cells.Methods：The eukaryotic expression plasmid pIRES2-EGFP-SLC22A18 was constructed and introduced by Lipofectamine 2000 into cultured U251 cells.SLC22A18 mRNA and protein expression was detected by RT-PCR and Western blot assay.U251 cells were divided into 4 groups：control group,transfected group,radiation group and combined treatmemt group.The number of colonies was assessed using a clonogenic assay.The cell growth inhibitition and apoptosis was assessed by MTT and flow cytometry.The tumor growth inhibition effect was further studied in vivo.The pIRES2-SLC22A18 was injected iniratumorally into established subcutaneous U251 glioma in nude mice mediated by Lipofectamine 2000.Results：The transfection of SLC22A18 gene into U251 cells was confirmed by RT-PCR and Western blot assay.The number of colonies was assessed using a clonogenic assay in the transfected group（60.6 ± 5.2）.That induced by radiation was decreased [（30.0 ± 3.6）,（13.0 ± 3.0）,（4.0 ± 1.0） ]with the increase of radiation doses（3,6,9 Gy）.MTT showed that SLC22A18 gene by itself induced strong inhibition effect on the growth of U251 cells [inhibition rate,IR（80.12 ± 5.75） ].The killing effect of radiation by itself on U251 cells was not strong [IR（17.05 ± 4.24） %,（17.34 ± 1.62） %,（18.71 ± 4.59） %]and increased with the increase of radiation doses（3,6,9 Gy）.When combined treatment of SLC22A18 gene transfection and radiation was used,that was significantly increased [IR（81.45 ± 5.32） %,（90.45 ± 1.65） %,（92.62 ± 2.12） %].The apoptotic rate of U251 cells induced by SLC22A18 gene transfection was 17.68%.That induced by radiation was increased（4.64%,4.87%,5.42%） with the increase of radiation doses（3,6,9 Gy）.The apoptotic rate was also significantly increased（18.42%,21.48%,23.92%） after combined treatment of SLC22A18 and radiation with different doses（3,6,9 Gy）.The antitumor enhancem

关 键 词：胶质瘤 基因 SLC22A18 转染 放疗敏感性 

分 类 号：R739.41[医药卫生—肿瘤] R730.55[医药卫生—临床医学]