^12C^6＋重离子诱导可溶性TRAIL表达增强及其抗肿瘤作用的实验研究  

Study on enhanced expression and anti-tumor effect of soluble TRAIL induced by ^12C^6＋ heavy ion irradiation

作　　者：田梅[1] 赵宝锋[1] 阮建磊[1] 李文建[2] 苏旭[1] 

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所,北京100088 [2]中国科学院兰州近代物理研究所

出　　处：《中华放射医学与防护杂志》2007年第3期238-241,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：兰州重离子加速器国家实验室资助项目;中国博士后基金资助项目（2005038364）

摘　　要：目的构建可被辐射诱导激活的表达载体pEgr-sTRAIL,研究其体外瞬时转染联合不同剂量的^12C^6＋离子辐照对人宫颈癌HeLa细胞凋亡和增殖活性的影响。方法SOEing PCR法克隆可溶性TRAIL基因,测序正确后连入pcDNA3.1-Egr质粒,构建pEgr-sTRAIL表达载体;采用GeneCompanionTM聚阳离子转染试剂体外瞬时转染人宫颈癌HeLa细胞;接受不同剂量的^12C^6＋照射后,RT-PCR法检测目的基因 mRNA水平的表达,流式细胞术检测细胞凋亡,克隆形成实验检测细胞增殖存活。结果成功构建了辐射诱导表达载体pEgr-sTRAIL;瞬时转染联合^12C^6＋离子照射可在mRNA水平上诱导sTRAIL表达增强;随照射剂量的增加,转染pEgr-sTRAIL的HeLa细胞凋亡率明显增加,克隆形成率明显下降。结论^12C^6＋离子可诱导重组质粒pEgr-sTRAIL在被转染的HeLa细胞中表达增高,体外基因-^12C^6＋离子联合治疗可诱导肿瘤细胞凋亡明显增多,具有显著的肿瘤抑制作用。Objective To construct pEgr-sTRAIL expression vector and investigate the apoptotic effect and survival level of its transient transfer on cervical cancer cell line HeLa followed by different doses of ^12C^6＋ heavy ion irradiation. Methods Human soluble TRAIL was cloned by SOEing method and then was linked with T-vector for sequencing. The correct fragment was inserted into pcDNA-Egr to construct pEgr-sTRAIL recombinant vector which can be activated and induced by ionizing irradiation. Then pEgr-sTRAIL recombinant vector was transferred into HeLa cells under mediation of GeneCompanionTM in vitro. After different doses of ^12C^6＋ heavy ion irradiation, the expression of TRAIL was detected by RT-PCR , the early apoptosis of transfected cells was measured by FACScan and the cell survival level was determined by colony formation assay. Results The expression vector pEgr-sTRAIL was successfully constructed. ^12C^6＋ heavy ion irradiation could enhance the expression of pEgr - sTRAIL at mRNA level. After irradiated by ^12C^6＋ heavy ion , the percentage of apoptotic cells in pEgr-sTRAIL transfected cells was significantly higher than that of non-transfected cells. The rate of colony formation of pEgr-sTRAIL transfected cells was significantly lower. Conclusion ^12C^6＋ heavy ion irradiation combined with pEgr-sTRAIL gene transfer can significandy induce the apoptosis of tumor cells and have strong anti-tumor effect in vitro.

关 键 词：pEgr-sTRAIL ^12C^6+离子 细胞凋亡 克隆形成 人宫颈癌HELA细胞 

分 类 号：R686[医药卫生—骨科学]