【摘要】 目的喉癌放疗不敏感的原因尚不清楚,有文献报道缺氧诱导因子(HIF)高表达与放疗不敏感有关。本研究探讨喉癌放疗不敏感与细胞缺氧的相关性。方法采用Hep-2乏氧放射模型,分别在常氧和5%O2低氧条件下培养Hep-2细胞6 h,然后给予4 Gy或8 Gy照射,在培养24 h和48 h后CCK8实验检测Hep-2细胞的生长活性变化。RT-PCR检测缺氧诱导Hep-2的mi R210表达。采用mi R210抑制物慢病毒载体转染Hep-2细胞,观察缺氧Hep-2细胞HIF1α的表达变化和辐射诱导细胞凋亡。结果缺氧细胞的细胞存活明显高于常氧组,缺氧6 h的Hep-2细胞mi R210表达明显高于正常培养的Hep-2细胞。转染mi R210抑制物慢病毒的Hep-2细胞HIF1α蛋白表达下调,辐射诱导的细胞凋亡率增加。结论低氧诱导的Hep-2细胞的辐射耐受与mi R210过表达有关。抑制mi R210表达可减少Hep-2细胞HIF1α的表达,增加喉癌放疗敏感性。更多还原

【Abstract】 Objective Hypoxia microenvironment has been associated with poor prognosis and resistance to radiotherapy of the laryngeal cancer. However, the mechanisms responsible for hypoxic survival of laryngeal cancer cells remain unclear. In this article, the correlation of radioresistance and hypoxia microenvironment in laryngeal cancer cells were explored and its regulation mechanism was investigated. Methods Radiotherapy cell model under hypoxia microenvironment was built by cultured Hep-2 cell under hypoxia condition for 6 h,and than given the cells with 4 Gy or 8 Gy radiation, and to detect cell viability by CCK8 assay after 24 h or 48 h. Real time q PCR was used to observed mi R210 expression in hypoxia condition and HIF1α expression changes and apoptosis induced by radiation were explored using mi R210 inhibitory vector transfection. Results After radiation, the cells survival of hypoxic cells was obviously higher than that of normal culture groups. mi R210 expression of Hep-2 cells after hypoxia treatment increased significantly. HIF1α protein level up-regulated under hypoxia treatment and decreased when cells were transfected with mi R210 inhibitory plasmid. And apoptosis rate of Hep-2cells with mi R210 inhibitory plasmid transfected increased than control plasmid group under hypoxia condition. Conclusion Radioresistance of Hep-2 induced by hypoxia is related with over-expression of mi R210. Inhibition of mi R210 expression can decrease the level of HIF1α in Hep-2 cells and increase the laryngeal cancer radiotherapy sensitivity.更多还原