摘要
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 investigate the role of transduction molecules and modulatory factors of signal pathways of Toll-like receptors （TLRs） in ovarian cancer. Methods：We collected peripheral blood mononuclear cell （PBMC） from 24 ovarian cancer patients,22 benign diseases, and 22 healthy females. Total RNA of the PBMCs was extracted, and reverse transcripted into cDNA. Expression levels of TLR1 ~TLR9 mRNA were then determined by real-time quantitative PCR. PBMCs were then either unstimulated （-） or stimulated with agonists for TLRs and assessed for IL-1β, IL-6,IL-12P40 by real-time PCR. Results：TLR1 ~TLR9 were all expressed in PBMC of the three groups and the expression levels of TLR2,TLR6 mRNA in patients with ovarian cancer were higher than the healthy controls （TLR2：F = 3.27 ,P 〈 0.05 ;TLR6：F = 2.21,P 〈 0.05）. Ovarian cancer patients aslo showed increased TLR2 levels compared to benign diseases group （F = 3.24,P 〈 0.05）.The expression of TLRs has no difference between benign diseases group and the healthy controls. After stimulated with HKLM for 24- h, ovarian cancer group demonstrated a significant up-regulation in expression of IL-1 β, IL-6,IL12-P40. The expressions of IL-1β, IL-6,IL12-P40 at the mRNA level were significantly higher in ovarian cancer group compared to healthy controls （F = 2.24, P 〈 0.05 ; F = 2.94, P 〈 0.05 ; F = 2.35, P 〈 0.05 ） and benign diseases group （F = 2.02,P 〈 0.05;F = 2.50,P 〈 0.05;F = 2.85,P 〈 0.05）. Furthermore,the level of IL-1β,IL-6,IL12-P40 mRNA had no difference between benign diseases group and healthy controls. Conclusion：TLR2 and TLR6 were highly expressed in ovarian cancer patients, inflammatory reaction mediated by IL-1β ,IL-6 ,IL-12P40 may be related to the growth and development of ovarian cancer.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2013年第3期308-313,共6页
Journal of Nanjing Medical University(Natural Sciences)
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)
国家临床检验重点专科建设单位
