LIU Zhonghua, HONG Chengjiao, ZHANG Baoguo. Heat shock transcription factor1 gene silenced with RNAi to enhance radio-sensitivity of cervical carcinoma cell[J]. Journal of Radiation Research and Radiation Processing, 2008,26(5):300-304.
LIU Zhonghua, HONG Chengjiao, ZHANG Baoguo. Heat shock transcription factor1 gene silenced with RNAi to enhance radio-sensitivity of cervical carcinoma cell[J]. Journal of Radiation Research and Radiation Processing, 2008,26(5):300-304.DOI:
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 transcription 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。
Abstract
In order to suppress HSF1 gene expression using RNA interference technique and explore the changes of the radiation sensitivity of cervical carcinoma cells, HSF1-pSilencer2.1-U6neo for siRNA expression was transfected into cervical carcinoma cells by lipofectamine. HSF1mRNA expression was detected by real time polymerase chain reaction and flow cytometry was employed to evaluate HSF1protein expression. In addiction, the radiation sensitivity of cervical carcinoma cells was assessed by clone forming assay. The results show that HSF1mRNA 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 HSF1gene 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.