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目的:观察卵巢癌患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中Toll样受体(Toll-likereceptors,TLRs)mRNA的表达水平,研究高表达TLRs在卵巢癌发病中的可能机制。方法:研究对象包括卵巢癌24例、妇科良性疾病22例及同期健康体检女性22例。采用密度梯度离心法分离PBMC后,提取总RNA,并逆转录为cDNA,采用实时荧光定量聚合酶链反应(real-time PCR)方法检测TLR1～TLR9 mRNA的表达水平;选择mRNA高表达的TLRs的相应激动剂刺激PBMC,采用Real-time PCR检测PBMC中前炎症细胞因子白细胞介素(IL)-1β、IL-6、IL-12P40的mRNA表达水平。结果:各组PBMC中TLR1～TLR9均有表达;卵巢癌组PBMC中TLR2、TLR6表达量显著高于健康对照组(TLR2:F=3.27,P<0.05;TLR6:F=2.21,P<0.05);卵巢癌组TLR2的表达量明显高于妇科良性疾病组(F=3.24,P<0.05);良性疾病组TLRs表达水平与健康对照组间无显著性差异;各组PBMC经TLR2的激动剂HKLM刺激24 h后,卵巢癌组IL-1β、IL-6、IL-12P40 mRNA表达明显增强,分别为健康对照组的2.24、2.94、2.35倍(P<0.05),为良性疾病组的2.02、2.50、2.85倍(P<0.05);良性疾病组与健康对照组比较无显著改变(F=1.11,F=1.18,F=0.82,P>0.05)。结论:卵巢癌患者PBMC中TLR2、TLR6表达水平显著增高,经其介导的炎症因子IL-1β、IL-6、IL-12P40 mRNA表达水平的改变可能在卵巢癌的发生发展中发挥作用。
OBJECTIVE: To observe the expression of Toll-like receptors (TLRs) in peripheral blood mononuclear cells (PBMC) of patients with ovarian cancer and to explore the possible mechanism of the high expression of TLRs in the pathogenesis of ovarian cancer. Methods: The study included 24 cases of ovarian cancer, 22 cases of benign gynecological diseases and 22 cases of women during the same period of physical examination. After isolation of PBMCs by density gradient centrifugation, the total RNA was extracted and reverse transcribed into cDNA. The expression of TLR1 ~ TLR9 mRNA was detected by real-time PCR. The high expression of TLRs PBMCs were stimulated with corresponding agonists, and the mRNA expression levels of pro-inflammatory cytokines IL-1β, IL-6 and IL-12P40 in PBMCs were detected by Real-time PCR. Results: The expression levels of TLR1 ~ TLR9 in PBMC of each group were significantly higher than those in healthy controls (TLR2: F = 3.27, P <0.05; TLR6: F = 2.21, P <0.05) . The expression of TLR2 in ovarian cancer group was significantly higher than that in benign gynecological disease group (F = 3.24, P <0.05). There was no significant difference between the expression of TLRs in benign disease group and healthy control group. The expression of TLR2 agonist HKLM The mRNA expression of IL-1β, IL-6 and IL-12P40 in ovarian cancer group was significantly increased after 24 h of stimulation, which were 2.24, 2.94 and 2.35 times respectively (P <0.05) in healthy group and 2.02 and 2.50 in benign group , 2.85-fold (P <0.05). There was no significant difference between benign disease group and healthy control group (F = 1.11, F = 1.18, F = 0.82, P> 0.05). CONCLUSION: The expression of TLR2 and TLR6 in PBMCs of patients with ovarian cancer is significantly increased. The changes of the expression of IL-1β, IL-6 and IL-12P mRNA may be involved in the pathogenesis of ovarian cancer.