摘要目的研究HAVCR2基因沉默对辐射诱发基因组不稳定肝细胞凋亡及细胞周期的影响.方法利用慢病毒介导的RNA干扰(RNAi)技术沉默辐射诱发基因组不稳定肝细胞中HAVCR2基因的表达,通过流式细胞术检测HAVCR2基因沉默对细胞周期及凋亡的影响;利用实时荧光定量PCR方法检测HAVCR2基因沉默后p53基因的表达变化.结果 慢病毒介导的RNAi技术能有效沉默HAVCR2基因的表达(=19.21,P＜0.05),与对照组相比,HAVCR2基因的沉默可导致基因组不稳定肝细胞G2期阻滞(t=-3.41,P＜0.05),细胞凋亡率降低(=3.65,P＜0.05);实时荧光定量PCR检测结果表明,HAVCR2基因的沉默诱发基因组不稳定肝细胞p53基因的表达下调(t=4.82,P＜0.05).结论HAVCR2 siRNA使辐射诱发基因组不稳定肝细胞凋亡率降低,使基因组不稳定肝细胞发生G2期的阻滞.

abstractsObjective To explore the effects of HAVCR2 siRNA on apoptosis and cell cycle in the radiation-caused genomic instable liver cells.Methods RNAi was used to inhibit HAVCR2 gene transcription.Cellular apoptosis and cell cycle distribution were detected by flow cytometry (FCM).Expression of p53 gene was assayed by real time fluorescence quantitative PCR.Results HAVCR2 gene was effectively inhibited by RNAi (t =19.21,P ＜ 0.05).After siRNA transferring,cell apoptosis (t =3.65,P ＜ 0.05) and p53 gene expression (t =4.82,P ＜ 0.05) were decreased,and G2-phase arrest was induced(t =-3.41,P ＜ 0.05).Conclusion HAVCR2 siRNA can decrease the generation of apoptosis in the genomic instable liver cells and blocks the cells at G2 phase.