MCP-1/CCR2信号轴在甲状腺乳头状癌合并桥本氏甲状腺炎患者中的表达及意义

目的 分析单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)及CC趋化因子受体2(CC chemokine receptor 2,CCR2)在人甲状腺乳头状癌(papillary thyroid carcinoma,PTC)合并桥本氏甲状腺炎(Hashimoto's thyroiditis,HT)的表达,以探讨MCP-1/CCR2信号轴在HT-PTC发展过程中与PTC发生、发展相关的生物学作用.方法 收集2013年6月至2014年6月在云南省第一人民医院乳腺及甲状腺肿瘤外科手术切除的人甲状腺新鲜组织(包括PTC组的癌组织、PTC-HT组的癌组织及病理证实的HT组的新鲜组织标本)及患者的外周血血清.运用实时荧光定量PCR、流式细胞微球芯片捕获技术(cytometric bead array,CBA)分析MCP-1/CCR2在人PTC、PTC-HT及HT组织和外周血血清的表达,表达谱基因芯片检测3组基因表达情况并进行多组差异基因筛选,构建共表达网络及显著性差异基因功能分析.结果 ①实时荧光定量PCR结果显示,CCR2 mRNA在HT甲状腺组织的表达水平(10.98 ±14.49)×10-3明显高于PTC癌组织(2.63 ±4.97)×10-3及PTC-HT癌组织[(3.77±4.65)×10“,x2=3.78,P=0.03];PTC患者的CCR2mRNA的表达水平与肿瘤直径呈负相关(P=0.01),与甲状腺球蛋白抗体(TGAb)及抗甲状腺过氧化物酶抗体(TPOAb)呈正相关(P =0.03,P=0.01);②CBA结果显示PTC-HT组患者外周血血清MCP-1的表达水平为(64.76±96.08) pg/mL,高于其他两组,但差异无统计学意义(F=1.04,P=0.36);③表达谱基因芯片结果显示,MCP-1 mRNA的表达水平在3组中差异无统计学意义(P＞0.05),CCR2 mRNA的表达在HT组较其他两组明显升高(P =0.014,FDR=0.082),共表达网络分析表明与CCR2表达相关的差异基因共有8个,显著相关性功能主要集中在与免疫相关的19项功能通路中.结论 MCP-1/CCR2信号轴在HT-PTC发展过程中与PTC的发生无协同互作相关性,CCR2可能通过与TGAb和TPOAb的正相关关系发挥作用.

Expression and significance of chemokine MCP-1 and its receptor CCR2 in patients with co-existing papillary thyroid cancer and Hashimoto's thyroiditis

Objective To investigate the expression of chemokine monocyte chemotactic protein-1 (MCP-1,also known as CCL2),and its receptor CC chemokine receptor 2 (CCR2) in the patients with coexisting papillary thyroid cancer (PTC) and Hashimoto's thyroiditis (HT),and to explore the biological effects of MCP-1/CCR2 signaling axis on the occurrence and development of PTC during the progression of HT-PTC.Methods Fresh thyroid tissues obtained by surgical removal (including thyroid carcinoma tissues from PTC and PTC-HT patients,and thyroid tissues of HT with pathological confirmation) and peripheral blood serum of above patients.The expression levels of CCR2 and MCP-1 in the thyroid tissues and blood samples were determined by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) and cytometric bead array (CBA).Multi-group differences in genetic screening,co-expression network construction and significant differences in gene function analyses were detected by cDNA microarray in the thyroid tissues from PTC,PTC-HT and HT patients.Results ①RT-PCR results showed that the mRNA expression levels of CCR2 were significantly higher in the HT thyroid tissue [(10.98 ± 14.49) x 10-3] than PTC [(2.63 ± 4.97) x 10-3] and PTC-HT carcinoma tissue [(3.77 ± 4.65) x 10-3,Chi square =3.78,P =0.03].The level in the PTC patients was negatively correlated with tumor diameter (P =0.01),and positively correlated with the levels of thyroglobulin antibody (TGAb) and anti-thyroid peroxidase antibody (TPOAb,P =0.03,P =0.01).②CBA results indicated that the serum MCP-1 level was higher in the PTC-HT patients (64.76 ± 96.08 pg/mL) than the other 2 groups,but without statistically significance (F =1.04,P =0.36).③Gene microarray displayed that there was no significant difference in the mRNA level of MCP-1 among the 3 groups (P ＞ 0.05),but that of CCR2 was obviously higher in the HT group than the other 2 groups (P =0.014,FDR =0.082).Co-expression network analysis showed the there were 8 genes related to the expressional differences of CCR2,whose functions were mainly involved in the 19 immune-related functions.Conclusion MCP-1/CCR2 signaling axis has no synergistic interaction or correlation with PTC occurrence during the progression of HT-PTC.CCR2 may exerts its positive effect through TGAb and TPOAb.