RNAi抑制人宫颈癌细胞HSF1基因表达增强细胞辐射敏感性的实验研究  

Heat shock transcription factorl gene silenced with RNAi to enhance radio-sensitivity of cervical carcinoma cell

作　　者：刘中华[1] 洪承皎[1] 张保国[1] 

机构地区：[1]苏州大学放射医学与公共卫生学院,苏州215123

出　　处：《辐射研究与辐射工艺学报》2008年第5期300-304,共5页Journal of Radiation Research and Radiation Processing

基　　金：国家自然科学基金(10275083)资助

摘　　要：通过RNA(Ribonucleic acid)干扰技术下调人宫颈癌细胞中热休克转录因子HSF1(Heat shock tran- scription factor1,HSF1)基因的表达,探讨沉默HSF1基因后,对宫颈癌细胞辐射敏感性的影响。将表达siRNA的HSF1-pSilencer2.1-U6 neo质粒表达载体,采用脂质体转染法转入宫颈癌细胞,用实时荧光定量PCR(Real time polymerase chain reaction,RT-PCR)检测HSF1mRNA表达水平,流式细胞仪检测蛋白质量,克隆法检测细胞的辐射敏感性。结果发现,与空白组细胞及阴性对照组细胞相比转染HSF1A-pSilencer2.1-U6neo质粒表达载体的细胞HSF1mRNA表达明显下降,同时Hela细胞的辐射敏感性也随着HSF1基因表达的下调而增高。表明siRNA质粒表达载体HSF1A-pSilencer2.1-U6neo转染细胞后可以降低HSF1的表达,可以增强宫颈癌细胞的辐射敏感性,具有良好的临床应用前景。In order to suppress HSF1 gene expression using RNA interference technique and explore the changes of the radiation sensitivity of cervical carcinoma cells, HSFl-pSilencer2.1-U6neo for siRNA expression was transfected into cervical carcinoma cells by lipofectamine. HSFlmRNA expression was detected by real time polymerase chain reaction and flow cytometry was employed to evaluate HSFlprotein expression. In addiction, the radiation sensitivity of cervical carcinoma cells was assessed by clone forming assay. The results show that HSFlmRNA expression of the cells transfected HSF1A-pSilencer2.1-U6neo descends obviously compared with control. Meanwhile the radiation sensitivity of Hela cells increases with the descension of HSF1 gene expression. Experiment has been confirmed that siRNA plasmid expression vector HSF1A-pSilencer2.1-U6neo successfully suppresses HSFlgene expression. Plasmid expression vector of HSF1 gene siRNA can be use to enhance the radio-sensitivity of cervical carcinoma cells and may be used as a powerfully adjunct method for conventional radiotherapy.

关 键 词：热休克转录因子 RNA干扰 热休克蛋白 辐射敏感性 

分 类 号：R811[医药卫生—放射医学] R811.5[医药卫生—临床医学]