NF-κB信号通路在放射线诱导的腺样囊性癌细胞凋亡中的实验研究  

Experimental study on the NF-κB signaling pathway in radio-induced apoptosis of adenoid cystic carcinoma cells

作　　者：刘哲[1] 张东升[1] 吴军楼[1] 黄圣运[1] 韩俊庆[2] 张世周[1] 刘桂军[1] 牟文丽[3] 张捷[3] 

机构地区：[1]山东大学附属省立医院口腔颌面外科,山东济南250021 [2]山东大学附属省立医院肿瘤研究治疗中心 [3]山东大学附属省立医院临床科研中心

出　　处：《临床口腔医学杂志》2012年第5期259-262,共4页Journal of Clinical Stomatology

基　　金：山东省科学技术发展计划(2006GG2202034)

摘　　要：目的:探讨NF-κB信号转导通路在放射线诱导的人唾液腺腺样囊性癌(ACC)-2细胞凋亡中的变化规律。方法:采用脂质体2000将pBabe-SR-IκBa质粒瞬时转染入ACC-2细胞,Western blot法检测转染后IκBa蛋白表达的变化,将转染后的ACC-2细胞进行6种剂量(0、2、4、6、8、10 Gy)的高能X线照射,在照射后不同时间点(1、3、6、10、24、48 h)应用免疫细胞化学检测ACC-2细胞NF-κB p65的表达情况,应用Image Pro-Plus图像分析软件测其细胞核平均灰度值,并用SPSS 16.0软件对数据进行方差分析。结果:与转染pBabe空白质粒组和未转染质粒组比较,转染pBabe-SR-IκBa质粒ACC-2细胞不仅表达内源性IκBa,同时也表达SR-IκBa;接受相同放射线剂量照射后3 h细胞核平均灰度值升高(P<0.05)。SR-IκBa质粒组中,不同剂量组的细胞核平均灰度值均低于0 Gy组;约在接受照射后6～10 h,细胞核平均灰度值达到最低值;相同时间点上,3 h组细胞细胞核平均灰度值随放射剂量增加而降低(P<0.05)。结论:转染突变的IκBa基因可特异性抑制NF-κB信号通路,经放射线照射诱导凋亡后,NF-κB p65蛋白的表达量具有一定的时间-剂量效应规律。Objective：To investigate the change pattern of NF-κB signal transduction pathway in radio-induced apoptosis of adenoid cystic carcinoma cells（ACC-2）.Method：The lipidosome infection protocol was used to transiently transfect the pBabe-SR-IκBa pasmid into ACC-2 cell.The change of expression of IκBa protein was detected by Western blotting.The transfected ACC-2 cells were irradiated by different doses of high energy X-ray（0、2、4、6、8、10Gy）.Expression of NF-κB p65 protein in ACC-2 cells was examinated by immunocyteochemistry after various time points（1、3、6、10、24、48 h）.To apply the Image Pro-Plus image analytical system to analyze the gray scale.Result：Compared to the transfected and non-transfected pBabe plasmid group,the ACC-2 cells for transfection of the pBabe-SR-IκBa plasmid expressed endogenous IκBa protein as well as SR-IκBa protein.The average nuclear gray scale of pBabe-SR-IκBa plasmid group increased according to the intergroup data analysis（P 0.05）.Intragroup data analysis of SR-IκBa group showed that the average nuclear gray scale of various dose groups were lower than that of 0 Gy group,and the datas reached the deepest valley at 6～10 h after irradiation.At the same time point,we disclosed at 3 h after irradiation,the average nuclear gray scale of ACC-2 cells decreased as the dose of irradiation increased（P 0.05）.Conclusion：The NF-κB signaling pathway was inhibited after transfection of mutated IκBa gene.The expression of NF-κB p65 possesses dose-time dependent effects in radio-induced apoptosis.

关 键 词：NF-ΚBP65 放射线 腺样囊性癌 质粒转染 

分 类 号：R739.87[医药卫生—肿瘤] Q786[医药卫生—临床医学]