华西口腔医学杂志

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信号转导蛋白P65与放疗诱导口腔鳞状细胞癌细胞凋亡的关系

马利1 张东升1 吴军楼1 韩俊庆2 张世周1 刘桂军1 牟文丽3 张捷3   

  1.  1.山东大学附属省立医院口腔颌面外科;2.肿瘤研究治疗中心; 3.临床科研中心, 济南250021
  • 收稿日期:2011-06-25 修回日期:2011-06-25 出版日期:2011-06-20 发布日期:2011-06-20
  • 通讯作者: 张东升,Tel:0531-85186950
  • 作者简介:马利(1983—),男,山东人,硕士
  • 基金资助:

    山东省科技攻关基金资助项目(2006GG20002046)

Relationship between P65 and radiotherapy-induced oral squamous cell carcinoma cell line apoptosis

MA Li1, ZHANG Dong-sheng1, WU Jun-lou1, HAN Jun-qing2, ZHANG Shi-zhou1, LIU Gui-jun1, MU Wen-li3, ZHANG Jie3   

  1. 1. Dept. of Oral and Maxillofacial Surgery, Provincial Hospital Affiliated to Shandong University, Jinan 250021,China; 2. Cancer Center, Provincial Hospital Affiliated to Shandong University, Jinan 250021, China; 3. Medical Research Center, Provincial Hospital Affiliated to Shandong University, Jinan 250021, China
  • Received:2011-06-25 Revised:2011-06-25 Online:2011-06-20 Published:2011-06-20
  • Contact: ZHANG Dong-sheng,Tel:0531-85186950

摘要:

目的探讨不同剂量的X射线对口腔鳞状细胞癌(OSCC)细胞中信号转导蛋白P65表达的影响以及P65与放疗诱导口腔鳞状细胞癌细胞凋亡的关系。方法人舌鳞癌Tca8113细胞复苏后,在37 ℃,5%CO2的孵箱中培养。共分对照组、2、4、6、8、10 Gy剂量组,待细胞长至对数生长期后,对细胞分别应用上述照射剂量一次性照射。照射后继续培养,并在照射后的不同时间点(1、3、6、10、24、48 h)应用免疫细胞化学和Western blot检测细胞中P65的表达量,并同时应用流式细胞仪和末端核苷酸转移酶介导的生物素化的dUTP末端标记(TUNEL)法检测不同X射线剂量组的细胞凋亡率。结果不同X射线剂量组的细胞浆内P65蛋白的表达量与对照组相比差异均具有统计学意义(P<0.05),而不同时间点组的细胞浆内P65蛋白表达量相互比较,3 h组的细胞浆内P65蛋白表达量与其他时间点组相比差异有显著性(P<0.05)。不同剂量组的凋亡率与对照组相比,差异均具有统计学意义(P<0.05),而不同时间点组的凋亡率相互比较时,3 h组的凋亡率与其他时间点组相比差异有统计学意义(P<0.05)。结论X射线可以激活口腔鳞状细胞癌细胞中核因子-κB(NF-κB) P65信号转导通路,而P65在细胞浆中表达与放疗诱导口腔鳞状细胞癌细胞凋亡具有正相关性,激活后进入细胞核内的P65蛋白可能具有抵抗细胞凋亡的作用。

关键词: P65, 口腔鳞状细胞癌, 细胞凋亡

Abstract:

Objective To investigate the effects of different X-ray doses on the expression of nuclear factor-κB (NF-κB) P65 in human oral squamous cell carcinoma cell(OSCC) line and the relationship between NF-κB P65 and radiation-induced OSCC cell line apoptosis. Methods The squamous cell carcinoma of Tca8113 cell was cultivated in the 37 ℃, 5%CO2 incubator after recovery. The experiment samples were divided into six groups(control group, 2, 4, 6, 8, 10 Gy). After growing to logarithm period, Tca8113 cells were irradiated using above-mentioned X-ray doses. The immunocyteochemistry and Western blot were used to detect the expression of NF-κB P65 after irradiation in various times(1, 3, 6, 10, 24, 48 h). The apoptosis rates under different radiotherapy dose were detected by flow cytometer and TDT-mediated dUTP-biotin nick end labeling(TUNEL). Results Compared with the control group, cytoplasm expression of P65 under different X-ray doses had statistically significant differences(P<0.05). While the cytoplasm P65 protein expression at different time were compared each other, the 3 h group demonstrated significant difference(P< 0.05). Apoptosis rates in various groups, compared with control group, had statistically significant differences(P<0.05). While the groups at different time points were compared each other, the apoptosis rates of 3 h group had significant differences(P<0.05). Conclusion X-ray can activate the NF-κB P65 in oral squmaous cell carcinoma cell lines. The correlation between expressional quantity of P65 and radiotherapy induced apoptosis of oral squamous cell carcinoma cell lines possesses positive correlation. The activated and intranuclear P65 may have radiotherapy resistant effect.

Key words: P65, oral squamous cell carcinoma, cell apoptosis