蛋白激酶C-δ在热诱导舌鳞癌Tca8113细胞凋亡中的作用研究  

Role of protein kinase C-δ in hyperthermia-induced apoptosis in tongue squamous cell carcinoma Tca8113 cells

作　　者：蒋文[1,2] 边莉[3] 李瑰琦[2] 马丽菊[3] 唐睿珠[3] 何永文[1] 

机构地区：[1]昆明医学院附属口腔医院口腔研究所,云南昆明650031 [2]荆州市第一人民医院口腔科,湖北荆州434000 [3]昆明医学院第一附属医院病理科,云南昆明650032

出　　处：《华西口腔医学杂志》2010年第5期539-542,546,共5页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目(30760272和30960422);教育部新世纪优秀人才支持计划基金资助项目(NCET-07-0388);云南省自然科学基金资助项目(2006C0038Q和2009CC021)

摘　　要：目的探讨热诱导舌鳞癌Tca8113细胞凋亡过程中蛋白激酶C-δ(PKC-δ)的作用。方法应用PKC-δ活化抑制剂Rottlerin和等体积的Rottlerin溶剂二甲基亚砜(DMSO)分别预处理Tca8113细胞30 min后,43℃水浴法热诱导Tca8113细胞0、40、80、120 min,流式细胞仪检测细胞凋亡率、线粒体膜电位强度变化,酶标仪比色法检测Cas-pase-3活性,Western杂交分析PKC-δ的活化。结果 Rottlerin抑制热诱导的PKC-δ裂解活化;抑制热诱导Tca8113细胞凋亡、降低线粒体膜电位及Caspase-3活性。相同40、80、120 min加热时间,经Rottlerin与未经Rottlerin预处理的Tca8113细胞相比,两者凋亡率、线粒体膜电位及Caspase-3活性在统计学上有显著性差异(P<0.01)。结论活化的PKC-δ可促进热诱导Tca8113细胞凋亡,PKC-δ裂解活化是热诱导Tca8113细胞凋亡的机制之一。Objective To study the role of protein kinase C-δ（PKC-δ） in hyperthermia-induced apoptosis in human tongue squamous cell carcinoma Tca8113 cells.Methods Tca8113 cells were treated at 43 ℃ in a heating water bath for 0,40,80,120 min after pretreatment with Rottlerin,a specific inhibitor of PKC-δ,and equal volume dimethyl sulfoxide（DMSO） for 30 min,respectively.The cells were stained by propidium iodide（PI） and Rhodamine 123 to analysis apoptotic rate and the changes of mitochondrial transmembrane potential by flow cytometry（FCM）.The total proteins were extracted for Western blotting analysis of activation and proteolysis of PKC-δ,and for colorimetric assay of relative activity of Caspase-3.Results Hyperthermia could induce proteolysis and activation of PKC-δ,and this was attenuated by Rottlerin.Apoptotic rate,decreasing of mitochondrial transmembrane potential and activity of Caspase-3 which being induced by hyperthermia in Tca8113 cells were inhibited by PKC-δ specific inhibitor Rottlerin.There were significantly statistical differences in apoptosis rates,mitochondrial transmembrane potential and activity of Caspase-3 between Rottlerin-and non-Rottlerin-pretreated cells after hyperthermia for 40,80,120 min（P 0.01）.Conclusion Activated PKC-δ may facilitate hyperthermia-induced apoptosis in Tca8113 cells,and may be one of the mechanisms of apoptosis induced by hyperthermia.

关 键 词：舌鳞状细胞癌 蛋白激酶C-δ 凋亡 热疗 

分 类 号：R739.86[医药卫生—肿瘤]