内源性NO在人宫颈癌Hela细胞株合成对顺铂敏感性的影响  

The influence of endogenous NO (induced by INF-γ) to the cisplatin sensitivity of human cervical carcinoma cell line Hela

作　　者：房姝妍[1] 张青 张瑞萍 侯杰 张锦阳 高志安 

机构地区：[1]辽宁医学院辽河油田总医院研究生培养基地,辽宁盘锦124000 [2]辽河油田总医院妇婴医院,辽宁盘锦124000 [3]辽河油田总医院石化医院,辽宁盘锦124000 [4]辽宁医学院附属第一医院病理科,辽宁锦州121000

出　　处：《中外医疗》2012年第27期20-22,24,共4页China & Foreign Medical Treatment

摘　　要：目的通过细胞因子γ-干扰素(INF-γ)诱导内源性一氧化氮(Nitric Oxide,NO)的产生,探讨其对人宫颈癌Hela细胞株顺铂(DDP)敏感性的影响。方法用NO测试盒检测一定浓度的INF-γ处理人宫颈癌Hela细胞在不同时间NO的产生量及一定作用时间下不同浓度的INF-γ处理Hela细胞NO的产生量;采用四甲基偶氮唑蓝(MTT)法检测经一定浓度及一定时间INF-γ处理后的人宫颈癌Hela细胞在不同浓度顺铂作用后的生存率;分别加入一氧化氮合酶(Nitric Oxide Synthase,NOS)的抑制剂NG-甲基-L-精氨酸(L-NMMA)及NO的合成原料L-精氨酸(L-arginine,L-Arg)再次以MTT法检测Hela细胞生存率的变化。结果体外培养的Hela细胞在经INF-γ作用后第8小时NO生成量达高峰,随后呈下降趋势,而不加INF-γ的空白对照组,Hela细胞NO的生成量无明显变化(P<0.05);Hela细胞NO的生成量与INF-γ的浓度之间有一定的依赖性。采取MTT法检测:经1nMINF-γ作用8 h的实验组Hela细胞与不加INF-γ的对照组相比,低浓度DDP作用时前者Hela细胞生存率显著下降(P<0.05),但随DDP浓度升高后两组的生存率无显著差异;经1nMINF-γ处理8 h的Hela细胞加入L-NMMA组Hela细胞在DDP作用后的生存率显著提高(P<0.05),而加入L-Arg组结果则与之相反。结论人宫颈癌Hela细胞在一定剂量、一定时间INF-γ的作用下,能够诱导iNOS产生内源性NO,且NO产生的量与INF-γ存在时间、剂量依赖关系;经INF-γ诱导产生的内源性NO可提高人宫颈癌Hela细胞株对DDP的敏感性。Objective To investigate the influence for the sensitivity to DDP in human cervical cancer Hela cell line through inducing endogenous NO by INF-γ.Methods Use NO Assay Kit to detect the production of NO under different groups of time when Hela cells are affected by INF-y and concentration of INF-γ.Employ MTT to detect the survival rate of human cervical cancer Hela cells which are treated by INF-γ at certain concentration and time among different concentration of DDP.L-NMMA which is an inhibitor of NOS and L-Arg which is a raw material of NO should be added respectively to detect changes of survival rates of Hela cells again by using MTT.Results The production of NO which is produced by the process that Hela cells are treated by INF-γ reaches the peak at the eighth hour,and then it decreases.However,the production of NO does not show obvious changes（P0.05） in control group which INF-γ is not added to.The production of NO in Hela cells depends on the concentration of INF-γ to some extent.Use MTT to carry out an inspection.By comparing control groups where Hela cells have been treated by 1nMINF-γ for 8 hours with experiment groups where INF-γ is not added,it is known that the survival rate of Hela cells in experiment groups decreases remarkably when the concentration of DDP is low.However,the survival rate of Hela cells in these two groups does not show any obvious difference.The survival rate of Hela cells treated by DDP increases significantly when L-NMMA is added to Hela cells treated by 1nMINF-γ for 8 hours（P0.05）.On the other hand,the survival rate of Hela cells where L-ARG is added decreases significantly（P0.05）.Conclusion Incubated with INF-γ under proper concentration and proper time,Hela cells can produce endogenous NO,there are an depending on relationship between the quantity of endogenous NO and time,dosage of INF-γ;endogenous NO can increase chemosensitivity to DDP.

关 键 词：宫颈癌 一氧化氮 顺铂 Γ-干扰素 化学敏感性 

分 类 号：R737[医药卫生—肿瘤]