TLR4受体在肝癌细胞株HepG-2表达.docVIP

TLR4受体在肝癌细胞株HepG-2表达[摘要] 目的：探讨TLR4受体在肝癌细胞株HepG-2和肝癌组织中的表达。方法：用RT-PCR和Western-blot方法检测肝癌细胞株HepG-2中TLR4基因的转录和蛋白的表达的情况，同时用免疫组化的方法检测肝癌组织中TLR4的表达的情况。结果：肝癌细胞株HepG-2中存在TLR4基因的转录和蛋白的表达。TLR4受体主要表达位于肝癌组织细胞的细胞膜上，其阳性率约为76.7%。结论：肝癌细胞株HepG-2和肝癌组织中均有TLR4的表达。 [关键词] HepG-2；肝癌；TLR4；RT-PCR；Western-blot [中图分类号] R73-3[文献标识码]A [文章编号]1673-7210（2008）06（c）-019-02 The expression of TLR4 on human hepatocellular carcinoma cells HepG-2 SUI Chun-yang1, LI Jia1, HU Yong2, GUO Ren-xuan3 (1.Department of General Surgery, Fengtian Hospital, Shenyang Medical University, Shenyang 110001, China;2.the Second People’s Hospital of Panjin City,Panjin124010,China;3.Department of General Surgery, the First Affiliated Hospital of China Medical University, Shenyang 110001, China) [Abstract] Objective:To investigate the expression of TLR4 on human hepatocellular carcinoma cells HepG-2 and tissue. Methods:To detect the expression of TLR4 mRNA and protein on human hepatocellular carcinoma cells HepG-2 by RT-PCR and Western-blot. To detect the expression of TLR4 protein on human hepatocellular carcinoma tissue by S-P immunohistochemical method. Results: The TLR4 could be detected on human hepatocellular carcinoma cells HepG-2.Cells staining for TLR4 show yellow or brow granules located on cell membrane in human hepatocellular carcinoma tissue. The positive rate was 76.7%.Conclusion:Human hepatocellular carcinoma cells HepG-2 and cells of human hepatocellular carcinoma tissues have the expression of TLR4. [Key words] TLR4；Hepatocellular carcinoma；HepG-2；RT-PCR；Western blot 研究表明，TLR4不仅表达在免疫系统相关的细胞中，许多肿瘤细胞表面都表达TLR4，这可能与肿瘤的免疫微环境的建立与维持有关[1]。例如胃癌的细胞表面有TLR4的表达，其使胃癌细胞与幽门螺杆菌相互作用而诱导产生促胃癌细胞生长的因子[2]。TLR4在一些肝脏疾病如病毒性肝炎、肝硬化、肝纤维化、酒精性肝病等的发生及发展机制中亦有重要作用 [3]。那么，TLR4在肝癌细胞中是否有表达，是否影响肝癌的发生发展呢？本实验将检测肝癌组织和细胞株HepG-2是否有TLR4的表达。 1材料与方法 1.1 细胞培养 HepG-2（购自肿瘤研究所）在含10%胎牛血清的（FBS）的RPMI-1640培养基中常规培养，95%湿度，5% CO2，37℃。每2~3天换培养基1次，每周传代1次。选生长状态好，处于对数生长期的细胞进行试验。 1.2 RT-PCR 取1×107细胞，加入Trizol 1 ml,按总RNA提取试剂盒(Invitro