TLR3激动剂poly(I:C)对胃癌BGC-823细胞生物学特征的影响  

The effects of TLR3 agonist poly(I:C) on human gastric adenocarcinoma cell line BGC-823 features

作　　者：屈静[1] 张建[1] 

机构地区：[1]山东大学药学院免疫药理与免疫治疗学研究所,济南250012

出　　处：《中国免疫学杂志》2012年第9期792-796,806,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(81172789;30972692)资助项目

摘　　要：目的:poly(I:C)是双链RNA(dsRNA)的结构类似物,可被模式识别受体TLR3、RLRs识别,具有抗肿瘤作用。本文旨在研究poly(I:C)对胃癌BGC-823细胞生物学特征的影响并进行机制探讨。方法:以胃腺癌细胞系BGC-823细胞为模型,采用半定量RT-PCR检测TLR1-TLR10 mRNA基础表达水平,然后用MTT法、CCK-8法、EdU掺入法检测poly(I:C)对BGC-823细胞的增殖能力的影响;MTT法、CFSE-7-AAD法评价NK细胞对poly(I:C)处理后的BGC-823细胞杀伤能力的影响;实时定量PCR法检测细胞因子mRNA水平的变化;流式细胞术分析BGC-823细胞中TLR3、Cyclin D1表达水平以及NKL细胞中CD107a、Perforin和TNF-α的表达水平。结果:TLR3在BGC-823细胞有明显表达。poly(I:C)可抑制BGC-823细胞的增殖,这一过程的主要分子特征是DNA合成减弱、Cyclin D1的表达水平降低、炎症因子的mRNA表达水平下降;同时NK细胞对于poly(I:C)处理后的BGC-823细胞的杀伤功能增强,NK细胞表达CD107a、Perforin、TNF-α的能力提高。结论:poly(I:C)可直接抑制肿瘤的生长并提高BGC-823细胞对NK细胞杀伤的敏感性,为设计和利用基于TLR的抗肿瘤治疗药物提供有益的启示。Objective：Polyinosinic-polycytidylic acid（poly（I：C）） is a synthetic analog of double-stranded RNA,which can be recognized by Toll-like receptor 3（TLR3） and retinoic acid inducible gene I（RIG-I）-like receptors（RLRs）,and exerted anti-cancer effect.However,the effect of poly（I：C） on gastric cancer have not been well explored.Our research aims to demonstrate the effect of poly（I：C） on human gastric adenocarcinoma cell line BGC-823 and investigate related molecular mechanisms.Methods：The basic mRNA levels of TLR1-TLR10 in BGC-823 cells were determined by semi-quantitative RT-PCR.After treated with poly（I：C） for 24 h,the proliferation rates of BGC-823 cells were measured by MTT assay,CCK-8 assay and EdU incorporation assay,and NK cell cytolysis activity against BGC-823 cells were analyzed by MTT assay,CFSE-7-AAD assay.Cellular total RNA was extracted,and cytokine mRNAs were detected by real-time PCR.Flow cytometry was used for analysis of the expression of TLR3 and Cyclin D1 on BGC-823 cells and the level of CD107a,Perforin and TNF-α in NK cells.Results：TLR3 is highly expressed in BGC-823 cells.When treated with poly（I：C）,the proliferation rate of BGC-823 cells were inhibited,accompanied with the decreased level of DNA replication,Cyclin D1 and the mRNA levels of several cytokines;Meanwhile,the cytolysis activity of NK cell was increased,concomitant with increased secretion of CD107a,Perforin and TNF-α.Conclusion：poly（I：C） can directly suppress the proliferation of BGC-823 cells and enhance the sensitivity of BGC-823 cell to NK cell cytolysis,which suggests a new approach based on TLRs for anti-tumor therapeutic application.

关 键 词：poly(I:C) BGC-823细胞 TLR3 NK细胞 

分 类 号：R735.2[医药卫生—肿瘤]