中华放射医学与防护杂志

田梅,赵宝锋,阮建磊,等.¹²C⁶⁺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[J].中华放射医学与防护杂志,2007,27(3):238-240.TIAN Mei,ZHAO Bao-feng,RUAN Jian-lei,et al.Study on enhanced expression and anti-tumor effect of soluble TRAIL induced by ¹² C⁶⁺ heavy ion irradiation[J].Chin J Radiol Med Prot,2007,27(3):238-240

¹²C⁶⁺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 ¹² C⁶⁺ heavy ion irradiation

投稿时间：2006-07-31

DOI：

中文关键词: pEgr-sTRAIL ¹²C⁶⁺离子细胞凋亡克隆形成人宫颈癌HeLa细胞

英文关键词:pEgr-sTRAIL ¹²C⁶⁺ heavy ion Apoptosis Colony formation HeLa

基金项目: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)

作者	单位	E-mail
田梅	100088 北京, 中国疾病预防控制中心辐射防护与核安全医学所
赵宝锋	100088 北京, 中国疾病预防控制中心辐射防护与核安全医学所
阮建磊	100088 北京, 中国疾病预防控制中心辐射防护与核安全医学所
李文建	中国科学院兰州近代物理研究所
苏旭	100088 北京, 中国疾病预防控制中心辐射防护与核安全医学所	suxu@nirp.cn

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中文摘要:

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¹²C⁶⁺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GeneCompanion^TM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¹²C⁶⁺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¹²C⁶⁺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 ¹²C⁶⁺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-¹²C⁶⁺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。

英文摘要:

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 ¹²C⁶⁺ 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 GeneCompanion^TM in vitro. After different doses of ¹²C⁶⁺ 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. ¹²C⁶⁺ heavy ion irradiation could enhance the expression of pEgr - sTRAIL at mRNA level. After irradiated by ¹²C⁶⁺ 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 ¹²C⁶⁺ heavy ion irradiation combined with pEgr-sTRAIL gene transfer can significantly induce the apoptosis of tumor cells and have strong anti-tumor effect in vitro.

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