肿瘤坏死因子-α对人喉鳞癌细胞放疗增敏作用的实验研究  

Experimental study on effect of TNF-α on reversing radioresistance in laryngeal squamous carcinoma cell

作　　者：朱圣明[1] 宋仕茂[1] 吴峰[1] 骆志国[1] 丁珺[1] 吴俊波[1] 

机构地区：[1]湖北省十堰市郧阳医学院附属太和医院肿瘤科,湖北十堰442000

出　　处：《重庆医学》2010年第2期133-136,共4页Chongqing medicine

基　　金：湖北省卫生厅重点资助项目(批准号:200ZJX3A20)

摘　　要：目的探讨人肿瘤坏死因子α(TNF-α)对放射抗拒性人喉鳞癌细胞增殖、凋亡的影响及对其放射抗拒性的逆转作用。方法(1)利用MTT法检测TNF-α对人喉鳞癌放射抗拒性细胞系Hep-2R增殖的影响;(2)TUNEL法和流式细胞仪检测TNF-α对Hep-2R细胞凋亡的影响;(3)RT-PCR法测定TNF-α处理、TNF-α联合放疗处理Hep-2R后,细胞凋亡相关基因BaxαmRNA的表达水平;(4)克隆形成实验检测TNF-α对Hep-2R细胞放射敏感性的逆转作用;(5)移植瘤实验检测TNF-α对Hep-2R细胞放射敏感性的逆转。结果(1)TNF-α对人喉鳞癌放射抗拒性细胞系Hep-2R的生长增殖具有明显的抑制作用,且具有浓度依赖性,其中400u/mL为理想药物浓度。(2)TUNEL结果显示:TNF-α处理后,Hep-2R细胞凋亡率明显增加;流式细胞分析显示:对照组、TNF-α处理组和TNF-α联合放疗处理组Hep-2R细胞的凋亡率分别为10%,30%和55%,差异具有统计学意义(P<0.05)。(3)RT-PCR检测结果表明,TNF-α处理组和TNF-α联合放疗处理组Hep-2R细胞的Baxα基因表达均较对照组细胞明显上调。(4)克隆形成实验结果显示:对照组、TNF-α处理组、放疗组和TNF-α联合放疗处理组细胞的克隆形成率分别为100%,(80±3)%,(58±2)%和(39±2)%,差异有统计学意义(P<0.05)。(5)裸鼠移植瘤实验结果显示:TNF-α处理组,放疗组和TNF-α联合放疗处理组细胞抑瘤率分别为(15.2±1.9)%,(35.4±2.1)%和(55.1±2.9)%,三者比较差异有统计学意义(P<0.05)。结论TNF-α可能通过抑制人喉癌细胞增殖,促进细胞凋亡,从而起到有效的放疗增敏作用。Objective To investigate the effects of TNF-α on the proliferation,apoptosis and reversing radioresistance in laryn geal squamous carcinoma cells. Methods （1） The proliferation impacts by TNF-α on the radioresistant laryngeal squamous carcinoma cell line Hep-2R,which was established in previous study,was determined by MTT method; （2） TUNEL assay and flow cytometry were performed to investigate the apoptotic rates of Hep-2R cells; （3） Real-time polymerase chain reaction （RT-PCR） was used to detect the expression level of apoptosis associated gene baxa mRNA in Hep-2R cells treated with TNF-α, TNF-α-radiation combination respectively and in their blank control; （4） Clone formation assay was carried out to investigate the radiosensitivity changes in Hep-2R cells when TNF-α was used; （5） Transplantation tumor experiment in nude mouse was employed to determine the impacts on radiosensitivity of Hep-2R cells by TNF-α. Results （1） TNF-α suppressed the proliferation of Hep-2R cells significantly and the effect was dependent on the drug concentration and 400U/mL was the preferential drug concentration. （2） The re suhs from TUNEL revealed that the apoptosis rates of Hep-2R cells treated by TNF-α alone and TNF-α radiation combination were increased significantly comparing with their blank control cells,and consistent with these results, the data from FCM displayed that the apoptosis index of the blank control cells, the TNF-α treating cells and the combination treating cells were 1.2±0.3,3.6±0.5 and 6.6±0.7,respectively,and the corresponding apoptosis rates were 10%±1.3% ,30%±2.2% and 55%±2.7% ,respectively. These data from apoptosis rates and apoptosis index were different significantly among three group cells （P〈0.05）. （3） The expression of apoptosis associated gene bax was more up-regulation in Hep-2R cells treated by TNF-α alone and TNF-α radiation combination than in blank control cells. （4） Clone formation experiment results indicated the radiosensitivity

关 键 词：肿瘤坏死因子-Α 喉癌 增殖 凋亡 放疗增敏 

分 类 号：R739.65[医药卫生—肿瘤] R730.55[医药卫生—临床医学]