顺铂和氟尿嘧啶增强食管癌细胞NKG2D配体的表达及CIK细胞的杀伤活性

目的研究顺铂（Cisplatin,DDP）和氟尿嘧啶（5-fluorouracil,5-Fu）对人食管癌EC9706细胞NKG2D配体表达及CIK细胞杀伤活性的影响。方法 MTT法测定DDP、5-Fu的50%抑制浓度（IC50）;以1/2 IC50浓度DDP、5-Fu作用EC9706细胞72 h,RT-PCR检测DNA损伤修复系统相关信号分子的表达。流式细胞仪检测DDP、5-Fu作用前、后EC9706细胞NKG2D配体的表达。乳酸脱氢酶释放法检测效靶比20∶1时,CIK细胞对DDP、5-Fu作用前、后EC9706细胞的杀伤活性。结果 DDP、5-Fu的IC50分别为5μg/ml、10μg/ml...

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Bibliographic Details
Published in肿瘤防治研究 Vol. 39; no. 7; pp. 765 - 768
Main Author 李瑞君 梅家转 禹萌 刘桂举 冯睿婷 张晓娟
Format Journal Article
LanguageChinese
Published 2012
Subjects
NKG2D
氟尿嘧啶
细胞因子诱导的杀伤细胞
顺铂
食管癌
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Summary:目的研究顺铂（Cisplatin,DDP）和氟尿嘧啶（5-fluorouracil,5-Fu）对人食管癌EC9706细胞NKG2D配体表达及CIK细胞杀伤活性的影响。方法 MTT法测定DDP、5-Fu的50%抑制浓度（IC50）;以1/2 IC50浓度DDP、5-Fu作用EC9706细胞72 h,RT-PCR检测DNA损伤修复系统相关信号分子的表达。流式细胞仪检测DDP、5-Fu作用前、后EC9706细胞NKG2D配体的表达。乳酸脱氢酶释放法检测效靶比20∶1时,CIK细胞对DDP、5-Fu作用前、后EC9706细胞的杀伤活性。结果 DDP、5-Fu的IC50分别为5μg/ml、10μg/ml。DDP、5-Fu可上调DNA损伤修复系统相关信号分子mRNA的表达。DDP与EC9706细胞共孵育72 h后,EC9706细胞MICA、MICB、ULBP2、ULBP3表达较DDP作用前明显增强（P〈0.05）,ULBP1无明显变化（P〉0.05）。5-Fu与EC9706细胞共孵育72 h后,EC9706细胞MICA、ULBP2、ULBP3表达明显增强（P〈0.05）,MICB、ULBP1无明显变化（P〉0.05）。效靶比20∶1时,CIK细胞对EC9706细胞的杀伤活性为（37.08±0.62）%,CIK细胞对1/2 IC50DDP、1/2 IC505-Fu作用后的EC9706细胞杀伤活性分别为（59.33±2.10）%、（52.44±0.97）%,与作用前相比差异均有统计学意义（P〈0.05）。结论 DDP、5-Fu通过激活DNA损伤修复系统相关信号分子,提高EC9706细胞NKG2D配体的表达,从而增强EC9706细胞对CIK细胞杀伤的敏感度。
Bibliography:42-1241/R
Objective To explore the effects of Cisplatin（DDP） and 5-fluorouracil（5-Fu） on the expression of NKG2D ligands of human esophagus carcinoma cell EC9706 and cytokine-induced killer（CIK） cells cytotoxicity.Methods The IC50 of DDP,5-Fu against EC9706 cells were measured by MTT assay.Expressions of signal pathway molecules involved in DNA damage and repair system were detected by RT-PCR.The expression of NKG2D ligands（MICA,MICB,ULBP1,ULBP2,ULBP3） were analyzed by flow cytometery.Cytotoxicities of CIK cells against EC9706 cells before and after cultured by 1/2 IC50 DDP or 5-Fu were analyzed by LDH releasing assay at effector-to-target cell ratio（E∶T） of 20∶1.Results DDP,5-Fu could decrease the proliferation and survival rate of EC9706 cells,the IC50 was 5 μg/ml and 10 μg/ml,respectively,which increased mRNA expressions of signal pathway molecules involved in DNA damage and repair system.MICA,MICB,ULBP2,ULBP3 on EC9706 cells were over expressed after 72 h cultured with 1/2 IC50 DDP,while expression of MICA
ISSN:1000-8578