脂质体介导E1A-TK基因转染人肺腺癌细胞A549体外研究  

Study on E1A-TK Gene Transfected into Human Pulmonary Adenocarcinoma Cell A549 by Lipofectamine in vitro

作　　者：何祥梁 黄龙生 刘贵真 夏增献 郑广昆 杨小南 马中富[2] 何东华[3] 

机构地区：[1]广州经济技术开发区医院东区分院,广州510730 [2]中山大学附属第一医院普内科,广州510080 [3]中山大学达安基因临床检验中心,广州510080

出　　处：《热带医学杂志》2009年第1期15-18,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.30371628);广东省自然科学基金(No.5001777)

摘　　要：目的用脂质体转染真核表达质粒PcDNA3E1A-TK入人肺腺癌细胞A549,观察丙氧鸟苷(GCV)、X射线及二者联合对转染细胞的体外杀伤作用。方法用阳离子脂质体介导真核表达质粒PcDNA3E1A-TK转染人肺腺癌细胞A549,PCR、RT-PCR方法分别检测转染细胞中E1A-TK基因DNA整合、mRNA表达。MTT法测不同浓度GCV、不同剂量X射线及一定剂量X线照射后再给一定量GCV对转染细胞的生长抑制率。结果PCR、RT-PCR结果表明转染细胞有E1A-TK基因整合、mRNA表达。根据IC50,转染细胞对GCV、X射线的敏感性分别比亲代细胞提高111.3倍和2.9倍,在作用相同时间后,GCV+X线照射治疗对转染细胞的生长抑制作用比单纯GCV或X线治疗明显增强(P<0.01)。结论脂质体介导外源基因E1A-TK成功转入人肺腺癌A549细胞并获得表达;转染细胞获得了对GCV和X线的敏感性,GCV和X线对转染细胞有协同杀伤作用。Objective To transfect human puhnonary adenocarcinoma cells A549 with the eucaryotic expression plasmids pcDNA3E1A-TK by lipofectamine, and to determine the sensitivity of the transfected cells to the cytotoxie actions of ganeiclovir（GCV） and X-ray. Methods Eucaryotic expression plasmids containing E1A-TK gene were transfected into human pulmonary adenocarcinoma cells A549 by lipofeetamine. PCR and RT-PCR were used to examine E1A-TK gene DNA integration and mRNA expression in the transfected cells, respectively. GCV, X-ray or GCV ＋ X-ray induced growth inhibition of the transfeeted cells were determined by MTr cytotoxicity assay. Results E1A-TK gene DNA integration and mRNA expression were found in the transfected cells. The IC50 of GCV and X-ray on the transfected cells was increased 113.3 and 2.9 folds, respectively. GCV and X-ray acted synergistically to increase the cell death of the transfeeted cells. Conclusion E1A-TK gene was successfully transfected into human pulmonary adenocarcinoma cells A549 by lipofectamine. The gene was successfully expressed and conferred sensitivity of the transfeeted cells to GCV and X-ray. GCV plus X-ray exhibited synergistic killing effect on the transfeeted ceils.

关 键 词：E1A—TK基因 基因转染 基因表达 肺腺癌 放射敏感 

分 类 号：R734.2[医药卫生—肿瘤] R459.9[医药卫生—临床医学]