Lu Chien-Chang, Kuo Hsing-Chun, Wang Feng-Sheng, Jou Ming-Huey, Lee Ko-Chao, Chuang Jiin-Haur
Division of Colorectal Surgery, Department of Surgery, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung 833, Taiwan.
Department of Nursing, Chang Gung University of Science and Technology, Chiayi 61363, Taiwan.
Int J Mol Sci. 2014 Dec 24;16(1):159-77. doi: 10.3390/ijms16010159.
Toll-like receptors (TLRs) not only form an important part of the innate immune system but also serve to activate the adaptive immune system in response to cancer. Real-time PCR; immunohistochemical stain and Western blotting analyses were performed to clarify molecular alterations in colorectal cancer (CRC) patients. We identified Toll-like receptor 1 (TLR1), TLR2, TLR4 and TLR8 gene expression levels and downstream gene, i.e., interleukin-6 (IL-6), IL-8, interferon-α (IFN-α) and myeloid differentiation primary-response protein-88 (MyD88), expression levels in CRC patients and in cancer cell lines. CRC tissues have higher TLR1, TLR2, TLR4, TLR8, IL-6 and IL-8 gene expression levels than do the normal colon mucosa (p < 0.05). TLR2 expression varied in different cell types (mucosa and lymphocytes). There was no difference in the MyD88 and IFN-α gene expression levels between cancerous and normal colon mucosa. CRC patients had higher levels of IL-6 (p = 0.002) and IL-8 (p = 0.038) expression than healthy volunteers did; and higher IL-6 and IL-8 expression was also found to signify a higher risk of recurrence. CL075 (3M002) treatments can reduce the production of IL-8 in different cancer cell lines. The signaling pathway of TLRs in cancer tissue is different from that in normal cells; and is MyD88-independent. Higher expression levels of TLR1, TLR2, TLR 4 and TLR 8 mRNA were related to upregulation inflammatory cytokines IL-6 and IL-8 gene expression in tissue and to the upregulation of IL-6 in blood. The concentration of IL-6 in serum can be used as an indicator of the possibility of CRC recurrence. Treatment with 3M002 can reduce IL-6 production in vitro and may prevent CRC recurrence. Our findings provide evidence that TLR1, TLR2, TLR4 and TLR8 gene expression induce downstream IL-6 and IL-8 gene expression; detection of these expression levels can serve as a CRC marker.
Toll样受体（TLRs）不仅是先天性免疫系统的重要组成部分，而且在应对癌症时还能激活适应性免疫系统。进行了实时聚合酶链反应（PCR）、免疫组织化学染色和蛋白质印迹分析，以阐明结直肠癌（CRC）患者的分子改变。我们确定了Toll样受体1（TLR1）、TLR2、TLR4和TLR8基因的表达水平以及下游基因，即白细胞介素-6（IL-6）、IL-8、干扰素-α（IFN-α）和髓样分化初级反应蛋白88（MyD88）在CRC患者和癌细胞系中的表达水平。CRC组织中TLR1、TLR2、TLR4、TLR8、IL-6和IL-8基因的表达水平高于正常结肠黏膜（p<0.05）。TLR2的表达在不同细胞类型（黏膜和淋巴细胞）中有所不同。癌性和正常结肠黏膜之间MyD88和IFN-α基因的表达水平没有差异。CRC患者IL-6（p = 0.002）和IL-8（p = 0.038）的表达水平高于健康志愿者；并且还发现较高的IL-6和IL-8表达意味着复发风险更高。CL075（3M002）处理可降低不同癌细胞系中IL-8的产生。癌症组织中TLRs的信号通路与正常细胞不同；并且是MyD88非依赖性的。TLR1、TLR2、TLR 4和TLR 8 mRNA的较高表达水平与组织中炎性细胞因子IL-6和IL-8基因表达的上调以及血液中IL-6的上调有关。血清中IL-6的浓度可作为CRC复发可能性的指标。用3M002处理可在体外降低IL-6的产生，并可能预防CRC复发。我们的研究结果提供了证据，证明TLR1、TLR2、TLR4和TLR8基因表达可诱导下游IL-6和IL-8基因表达；检测这些表达水平可作为CRC的标志物。
