放疗对人唾液腺腺样囊性癌p65信号传导通路的影响

The effects of radiotherapy on p65 signal conduct access of adenoid cystic carcinoma cells

目的:研究放射治疗对人唾液腺腺样囊性癌(adenoidcysticcarcinoma,ACC)细胞核转录因子NF-κBp65(nucleartranscriptionfactor-κBp65)信号传导通路的影响,初步探讨放射线引起唾液腺腺样囊性癌p65信号传导通路变化的规律。方法:采用人唾液腺腺样囊性癌细胞系ACC-2进行细胞培养,在培养皿底部均匀贴附1～2层细胞时,予以不同剂量高能X线照射。继续进行细胞培养,观察细胞形态,并在不同时间段对细胞进行NFκBp65染色。采用LeicaQwinPlus细胞图像分析系统对染色后的图像进行灰度分析,并自动计算细胞核灰度值,计算其方差。通过比较不同照射剂量组及不同染色时间组之间腺样囊性癌细胞p65染色深度的差异,研究放射治疗对腺样囊性癌细胞p65信号传导通路的影响。对所得数据采用SPSS10.0统计软件包进行t检验。结果:高能X线照射后,贴壁细胞减少。各组测得图像灰度值与空白对照均有显著差异(P<0.05),各放疗组细胞核平均灰度值均低于空白对照组。高放射剂量组平均细胞核灰度值显著低于低放射剂量组(P<0.05)。除48h染色组外,各不同时间染色组平均细胞核灰度值均显著低于空白对照组(P<0.05)。48h染色组的平均细胞核灰度值与空白对照组比较无显著差异(P>0.05)。结论:高能X线可引起腺样囊性癌细胞p65信号传导通路开放。放射剂量与p65信号传导通路开放存在量效关系,即在一定范围内,随着放射剂量增加,p65信号通路的表达更为明显。p65信号传导通路开放具有时效性,即放射后p65信号传导通路开放增加,后逐渐减少,48h内恢复正常水平。

PURPOSE： To investigate the effects of radiotherapy on nuclear transcription factor--κB p65（NF-κB p65） of ACC-2 cells, and to find some regularities that radiotherapy causes p65 signal conduct access of ACC-2 cells. METHODS： Adenoid cystic carcinoma cell line ACC-2 was cultured.The cells were radiated with various doses when there were 1 to 2 layers of cells on the bottom of petri dish. Cell culture was continued with observation of their morphology. The cells were stained with NF-κB p65 antibody at different times. Leica Qwin Plus image analysis system was adopted to analyse the gray scale of nuclei that had been stained, to calculate the gray scale of nuclei and the variance among the nuclei. The effects of radiotherapy on NF-κB p65 of ACC-2 were assessed by comparing the staining gray scales among various radiation doses and staining times.The data were analyzed with SPSS 10.0 software package for Student＇s t test. RESULTS： Fewer adenoid cystic carcinoma cells survived after radiation of high energy X- ray. There was significant difference in gray scale of nuclei between radiation group and the control group. The staining depth of the cells postradiotherapy was greater than that of the control group （P〈0.05）.The average nuclear gray scale of high irradiation dose was significantly lower than that of low irradiation dose （P〈0.05）. The average nuclear gray scale of various staining times was significantly lower than that of the control group except for 48-hour group（P〈0.05）. There was no significant difference in gray scale of nuclei between the 48-hour group and the control group （P〉0.05）.CONCLUSION： Radiotherapy can activate NF-κB p65 signal conduct access of ACC-2 cells.There is a dose-effect relationship between irradiation dose and the enabling of p65 signal conduct access of ACC-2. The expression of p65 signal conduct access of ACC-2 will active more greatly with the increase of radiation dose in some distance. There is also a time-dependent response in the enabling of p65 signal conduct access of ACC-2.The p65 signal conduct access of ACC-2 will be activate after radiation of high-energy X-ray, and decrease after a period of time,then decrease to normal level in 48 hours. Supported by Natural Science Foundation of Shandong Province （Grant No.12003C03）.