辐射诱导转录子RIGb cDNA对HeLa细胞增殖的抑制作用  

Inhibition of HeLa Cell Proliferation by Stably Transfected=Radiation-Induced Transcript RIGb cDNA

作　　者：孙志增[1] 徐勤枝[1] 隋建丽[1] 周平坤[1] 

机构地区：[1]北京放射医学研究所,北京100850

出　　处：《中国生物化学与分子生物学报》2005年第3期384-389,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家高技术研究发展计划(863计划;No.2004AA221160);国家自然科学基金(No.30270423)资助~~

摘　　要：核酸序列同源性分析和RT -PCR确定辐射诱导转录子RIGb是染色质重构基因CHD6表达的一个剪接转录子.Northern杂交结果表明,0 . 5Gyγ射线诱导RIGbmRNA表达增加,但4Gy大剂量照射对其表达无明显的影响.正常成人组织Northern杂交结果显示,RIGb基因在心脏、肝脏和睾丸中有高表达.细胞生长曲线分析结果表明,转染和稳定表达RIGb的人宫颈癌细胞(HeLa)的增殖生长受到明显抑制,细胞周期分析发现G1/S期阻滞.A radiation induced gene(RIGb) was cloned by using differential display technique from human lung HEL cell line.Sequence homology comparison has found that RIGb shares 99% homology with chromodomain helicase DNA binding protein 6(CHD6) which is involved in chromatin remodeling and RT-PCR analysis showed that RIGb actually was a splicing variant of CHD6.Furthermore,Northern blotting analysis revealed that following exposure to 0.5 Gy gamma radiation, RIBb mRNA level of HeLa cells enhanced significantly in time-dependent manner with effective at 4 hours,reaching maximal at 8 hours,and then declined,being lowest at 24 hour.Interestingly,higher dose at 4 Gy did not affect the mRNA levels.Multiple human tissue Northern blot assay showed that RIGb has substantial expression mainly distributed in heart,liver and testis.Flow cytometry results demonstrated that overexpression RIGb transcript induced by stable transfected HeLa cell with RIGb sense expression plasmid cause G-1/S arrest and inhibit proliferation of the cell line.The results suggested that up-regulation of RIGb gene expression was capable to postpone the progress of cell cycle from G-1 to S phase and therefore decrease the proliferation of tumor cells.

关 键 词：辐射诱导基因 基因表达 细胞周期 细胞增殖 

分 类 号：Q274[生物学—细胞生物学] Q25