XIAP反义寡核苷酸对hep-2细胞放射治疗的增效作用

目的:通过XIAP反义寡核苷酸转染hep-2细胞株,观察细胞增殖、凋亡及放疗反应。方法:体外培养人喉鳞状细胞癌hep-2细胞株,脂质体介导反义寡核苷酸转染,6h后4Gyγ射线照射。24h后RT-PCR检测XIAPmRNA表达,流式细胞仪检测蛋白水平及凋亡率,MTT检测细胞存活情况。结果:各浓度的反义寡核苷酸转染组hep-2细胞形态改变,MTT结果为细胞凋亡率随剂量增加而增高。800nmol/LXIAP反义寡核苷酸转染,使XIAPmRNA表达下调,蛋白表达下降,细胞凋亡率增加(P<0.05);4Gy照射后反义寡核苷酸转染组蛋白表达下降明显,较对照组细胞凋亡率增加,细胞存活率明显下降(P<0.05)。结论:XIAP反义寡核苷酸转染hep-2细胞株能够降低蛋白表达,促进细胞的凋亡并能提高对放疗的敏感性。

Antisense oligonucleotides targeting XIAP induce apoptosis and enhance radiotherapeutic activity against hep-2 cells in vitro

Objective:To investigate the down-regulation effect of antisense oligonucleotides(AS ODNs)targeting X-chromosome-linked inhibitors of apoptosis(XIAP)on hep-2 cells apoptosis and radiotherapy sensitivity in vitro.Method:G4 AS ODN was transfected into cultured hep-2 cells which received radiation 6 hours later.Twenty-four hours after radiation,cells were observed under inverted phase contrast microscope and fluorescence microscope.Apoptosis rate,cell viability,expression of XIAP mRNA and protein were tested.Result:In cultured hep-2 cells,G4 AS ODN down-regulated XIAP mRNA expression.Furthermore,Tthe protein expression of XIAP and the cell viability decreased too.In contrast to that,the scrambled control ODN caused minor XIAP loss and less cell inhibition.In addition,G4 AS ODNs activated Hep-2 cells after the radiation of 4 Gy Co60 ray.Conclusion:XIAP is a viable target for cancer therapy in human laryngeneal neoplasms.In cultured Hep-2 cells,AS ODN targeting XIAP can down-regulate protein expression of XIAP,induce cell apoptosis and enhance the radiotherapy sensitivity.