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目的 :观察TLR1(Toll-like receptors 1)、TLR2和TLR6在卵巢癌患者外周血单个核细胞(peripheral blood mononuclear cells,PBMC)亚群中的表达水平,初步探索TLR1、TLR2和TLR6在卵巢癌发生发展中的作用机制。方法 :收集13例卵巢癌患者、13例女性妇科良性疾病患者及13例同期健康体检女性EDTA抗凝外周全血,采用流式细胞仪分别检测3组单核细胞、CD4+T淋巴细胞、CD8+T淋巴细胞、B淋巴细胞中TLR1、TLR2和TLR6表达水平;TLR1、TLR2和TLR6相应配体(Pam3CSK4、HKLM、FSL-1)分别刺激3组PBMC,采用real-time PCR检测PBMC中前炎症细胞因子IL-1β、IL-6、IL-8、TNF-αm RNA水平。结果:3组细胞亚群中TLR1、TLR2和TLR6均有表达,且主要表达于单核细胞;卵巢癌组单核细胞中TLR1、TLR2、TLR6的表达率分别为69.13%、59.43%、52.99%,显著高于良性疾病组(34.34%、25.32%、15.21%)和健康对照组(36.31%、26.63%、16.43%),差异具有统计学意义(P<0.05);良性疾病组与健康对照组间无显著性差异;3组间CD4+T淋巴细胞、CD8+T淋巴细胞及B淋巴细胞中TLR1、TLR2、TLR6的表达率无显著性差异;3组经TLR2的配体HKLM刺激24 h后,卵巢癌组IL-1βm RNA表达水平显著高于良性疾病组和健康对照组(F=2.05,P<0.05;F=2.19,P<0.05),良性疾病组与健康对照组无显著差异;卵巢癌组IL-6m RNA表达水平显著高于良性疾病组和健康对照组(F=1.40,P<0.05;F=1.99,P<0.05),良性疾病组与健康对照组无显著差异;3组间IL-8及TNF-αm RNA表达水平无显著性差异。3组经TLR6配体FSL-1刺激24 h后,卵巢癌组IL-6 m RNA表达水平显著高于良性疾病组和健康对照组(F=1.30,P<0.05;F=1.69,P<0.05),良性疾病组与健康对照组间无显著性差异;3组间IL-1β、IL-8及TNF-αm RNA表达水平无显著性差异。TLR1配体Pam3CSK4刺激24 h后,3组间IL-1β、IL-6、IL-8及TNF-αm RNA表达水平无显著差异。结论:卵巢癌患者单核细胞中高表达的TLR1、TLR2、TLR6介导的前炎症细胞因子的上调,在卵巢癌炎症微环境的形成和维持中发挥重要作用。
OBJECTIVE: To investigate the expression of TLR1, TLR2 and TLR6 in the peripheral blood mononuclear cells (PBMC) subsets in ovarian cancer patients and to explore the possible role of TLR1, TLR2 and TLR6 in ovarian cancer The mechanism of development. Methods: Totally 13 ovarian cancer patients, 13 women with benign gynecological diseases and 13 healthy women with anticoagulant peripheral blood from healthy volunteers were enrolled in this study. Flow cytometry was used to detect the expression of monocytes, CD4 + T lymphocytes, CD8 (Pam3CSK4, HKLM, FSL-1) were used to stimulate PBMCs of three groups respectively, and the expression of TLR1, TLR2 and TLR6 in PBMCs, T lymphocytes and B lymphocytes were detected by real time PCR Cytokines IL-1β, IL-6, IL-8, TNF-αmRNA levels. Results: The expression of TLR1, TLR2 and TLR6 in three groups of cells was mainly expressed in monocytes. The expression rates of TLR1, TLR2 and TLR6 in mononuclear cells of ovarian cancer were 69.13%, 59.43% and 52.99%, respectively (34.34%, 25.32%, 15.21%) and healthy control group (36.31%, 26.63%, 16.43%, respectively), the difference was statistically significant (P <0.05); benign disease group and healthy control group There was no significant difference between the three groups; there was no significant difference in the expression rates of TLR1, TLR2 and TLR6 in CD4 + T lymphocytes, CD8 + T lymphocytes and B lymphocytes among the three groups; after stimulated by TLR2 ligand HKLM for 24 h , The expression level of IL-1βmRNA in ovarian cancer group was significantly higher than that in benign disease group and healthy control group (F = 2.05, P <0.05; F = 2.19, P <0.05); there was no significant difference between benign disease group and healthy control group; The expression of IL-6m RNA in cancer group was significantly higher than that in benign disease group and healthy control group (F = 1.40, P <0.05; F = 1.99, P <0.05); there was no significant difference between benign disease group and healthy control group IL-8 and TNF-αm RNA expression levels were not significantly different. The expression of IL-6 mRNA in ovarian cancer group was significantly higher than that in benign disease group and healthy control group (F = 1.30, P <0.05; F = 1.69, P <0.05) after stimulation with TLR6 ligand FSL-1 for 24 h ), There was no significant difference between benign disease group and healthy control group. The expression level of IL-1β, IL-8 and TNF-αmRNA in the three groups had no significant difference. There was no significant difference in the expression levels of IL-1β, IL-6, IL-8 and TNF-αmRNA among the three groups after TLR1 ligand Pam3CSK4 stimulated for 24 h. CONCLUSION: The upregulation of TLR1, TLR2 and TLR6-mediated proinflammatory cytokines in monocytes of ovarian cancer patients play an important role in the formation and maintenance of ovarian cancer microenvironment.