摘要
目的观察卵巢癌(OC)患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中Toll样受体1(Toll-like receptors 1,TLR1)、TLR2及TLR6信号通路活化后前炎症细胞因子分泌水平,研究其在OC发病中的可能机制。方法纳入OC患者、女性妇科良性疾病(BC)患者及同期体检健康女性(NC)各13例,用密度梯度离心法分离PBMC并用TLRs的相应配体刺激PBMC,用CBA免疫荧光法检测细胞中前炎症细胞因子白细胞介素-1β(IL-1β)、IL-6、IL-8、肿瘤坏死因子-α(TNF-α)的分泌水平;用western blot检测MyD88、TRAF6、TANK、NF-κB和P-NF-κB的表达水平。结果 OC组、BC组、NC组未加配体刺激前,PBMC中IL-1β、IL-6、IL-8、TNF-α分泌水平差异均无统计学意义(P>0.05)。与未经配体刺激PBMC相比,OC组PBMC经TLR1配体Pam3CSK4刺激24 h后,IL-1β分泌水平增高,差异具有统计学意义(t=-2.667,P<0.05);TLR2配体HKLM刺激24 h后,OC组IL-1β、TNF-α分泌水平亦增高,差异有统计学意义(t分别为-5.391和-3.564,P均<0.05);相较于未经配体刺激时,BC组、NC组的PBMC在相应TLRs配体刺激后,IL-1β、IL-6、IL-8、TNF-α分泌水平变化无统计学意义。OC患者PBMC经Pam3CSK4、HKLM、FSL-1刺激后,TLRs信号通路蛋白MyD88、TRAF6、TANK及P-NF-κB表达上调。结论 OC患者PBMC中TLR1、TLR2、TLR6经相应配体刺激后可活化TLR信号通路,并诱导前炎症细胞因子IL-1β、TNF-α分泌水平增加,在OC的发生发展中发挥作用。
Objective To investigate the levels of pro-inflammatory cytokines secreted by Toll-like receptor-1（TLR1）,-2（TLR2） and-6（TLR6） activated peripheral blood mononuclear cells（PBMCs） in the patients with ovarian cancer（OC）,and their possible roles in the pathogenesis of OC.Methods The blood samples were collected from 13 patients with OC,13 with benign gynecological diseases（BGD） and 13 healthy females（NC）,and their PBMCs were isolated with the density gradient centrifugation method.Then,the PBMCs were activated with TLR ligands for 24 h,and the levels of TNF-α,IL-8,IL-1β and IL-6 in PBMCs were determined by flow cytometry,and the levels of MyD88,TRAF6,TANK,NF-κB and P-NF-κB in PBMCs by Western blot.Results There were no significant difference among OC,BGD and NC groups for the levels of IL-1β,IL-6,IL-8 and TNF-α in PBMCs unactivated by TLR ligands.After the PBMCs from OC patients were activated by the TLR1 ligand-Pam3CSK4 for 24 h,the level of IL-1β in PBMCs increased significantly（t =-2.667,P 0.05）.Similarly,after the PBMCs from OC patients were activated by the TLR2 ligand HKLM for 24 h,the levels of IL-1β and TNF-α in PBMCs also increased significantly（t =-5.391 and-3.564,P 0.05）.However,after the PBMCs from BGD and NC groups were stimulated with Pam3CSK4,HKLM or FSL-1（TLR6 ligand）,the levels of IL-1β,IL-6,IL-8 and TNF-α in PBMCs showed no significant change.In addition,after the PBMCs from OC patients were stimulated with Pam3CSK4,HKLM or FSL-1,the expression levels of MyD88,TRAF6,TANK and P-NF-κB in PBMCs were up-regulated significantly.Conclusion TLR signaling pathways in PBMCs from OC patients could be activated by TLR ligands,which further led to the increase of IL-1β and TNF-α levels,indicating that TLR signaling pathways might play a crucial role in the pathogenesis of OC.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2013年第8期584-587,共4页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金(30901344
81272324)
江苏省实验诊断学重点实验室基金(XK201114)
