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Upregulation of TLRs and IL-6 as a marker in human colorectal cancer.

Lu CC, Kuo HC, Wang FS, Jou MH, Lee KC, Chuang JH - Int J Mol Sci (2014)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Division of Colorectal Surgery, Department of Surgery, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung 833, Taiwan. cclu999@gmail.com.

ABSTRACT
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.

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qRT-PCR of Toll-like receptor 1 (TLR1), TLR2, TLR4 and TLR8 gene expression in normal and cancerous tissue (*p < 0.05).
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ijms-16-00159-f001: qRT-PCR of Toll-like receptor 1 (TLR1), TLR2, TLR4 and TLR8 gene expression in normal and cancerous tissue (*p < 0.05).

Mentions: Colorectal cancer tissues have higher TLR1, TLR2, TLR4 and TLR8 (Figure 1) gene expression levels in general than do the normal colon mucosa from the same patient (p < 0.05). However, protein expression of TLRs examined in colorectal cancer tissues from patients showed unexpectedly differential results from gene expression assay. There were strong TLR1 and TLR8 immunoreactivities in the cancer cells and in some inflammatory cells in the mucosa. Normal and cancerous tissues showed a significant difference in TLR1 (81.2% and 38.8% of score 1 in normal and cancerous tissue, respectively; 18.7% and 61.1% of score 2 in normal and cancerous tissue, respectively; p = 0.015) and TLR8 (81.2% and 33.3% of score 1, as weak, in normal and cancerous tissue, respectively; 18.7% and 66.6% of score 2, as strong, in normal and cancerous tissue, respectively; p = 0.006) protein expressions (Figure 2; Table 1). TLR2 and TLR4 immunoreactivity was present mainly in the tumor-infiltrating inflammatory cells (lymphocytes), which were morphologically identical to monocytes/macrophages. A significant difference (75% and 33.3% of score 1 in normal and cancerous tissue, respectively; 18.7% and 66.6% of score 2 in normal and cancerous tissue, respectively; p = 0.018) of TLR2 protein expression between normal and cancerous tissues was shown in the lymphocytes (Table 1). In the mucosa, TLR2 protein expression had no difference in either score 1 (62.5% and 83.3% in normal and cancerous tissue, respectively) and score 2 (37.5% and 16.6% in normal and cancerous tissue, respectively) cells of normal and cancerous tissues. Similarly, the expression of TLR4 was mainly in the inflammatory cells of the normal mucosa rather than cancerous tissue (81.2% and 77.7% of score 1 in normal and cancerous tissue, respectively; 18.7% and 22.2% of score 2 in normal and cancerous tissue, respectively; p = 0.018; Figure 2; Table 1). However, TLR1 and TLR8 were highly expressed in CRC cells. The related gene expression of IL-6 and IL-8 (Figure 3) in cancerous tissues was higher than the normal colon mucosa from the same patient (p < 0.05). There was no difference in the MyD88 and IFN-α gene expression between cancerous and normal colon mucosa.


Upregulation of TLRs and IL-6 as a marker in human colorectal cancer.

Lu CC, Kuo HC, Wang FS, Jou MH, Lee KC, Chuang JH - Int J Mol Sci (2014)

qRT-PCR of Toll-like receptor 1 (TLR1), TLR2, TLR4 and TLR8 gene expression in normal and cancerous tissue (*p < 0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307241&req=5

ijms-16-00159-f001: qRT-PCR of Toll-like receptor 1 (TLR1), TLR2, TLR4 and TLR8 gene expression in normal and cancerous tissue (*p < 0.05).
Mentions: Colorectal cancer tissues have higher TLR1, TLR2, TLR4 and TLR8 (Figure 1) gene expression levels in general than do the normal colon mucosa from the same patient (p < 0.05). However, protein expression of TLRs examined in colorectal cancer tissues from patients showed unexpectedly differential results from gene expression assay. There were strong TLR1 and TLR8 immunoreactivities in the cancer cells and in some inflammatory cells in the mucosa. Normal and cancerous tissues showed a significant difference in TLR1 (81.2% and 38.8% of score 1 in normal and cancerous tissue, respectively; 18.7% and 61.1% of score 2 in normal and cancerous tissue, respectively; p = 0.015) and TLR8 (81.2% and 33.3% of score 1, as weak, in normal and cancerous tissue, respectively; 18.7% and 66.6% of score 2, as strong, in normal and cancerous tissue, respectively; p = 0.006) protein expressions (Figure 2; Table 1). TLR2 and TLR4 immunoreactivity was present mainly in the tumor-infiltrating inflammatory cells (lymphocytes), which were morphologically identical to monocytes/macrophages. A significant difference (75% and 33.3% of score 1 in normal and cancerous tissue, respectively; 18.7% and 66.6% of score 2 in normal and cancerous tissue, respectively; p = 0.018) of TLR2 protein expression between normal and cancerous tissues was shown in the lymphocytes (Table 1). In the mucosa, TLR2 protein expression had no difference in either score 1 (62.5% and 83.3% in normal and cancerous tissue, respectively) and score 2 (37.5% and 16.6% in normal and cancerous tissue, respectively) cells of normal and cancerous tissues. Similarly, the expression of TLR4 was mainly in the inflammatory cells of the normal mucosa rather than cancerous tissue (81.2% and 77.7% of score 1 in normal and cancerous tissue, respectively; 18.7% and 22.2% of score 2 in normal and cancerous tissue, respectively; p = 0.018; Figure 2; Table 1). However, TLR1 and TLR8 were highly expressed in CRC cells. The related gene expression of IL-6 and IL-8 (Figure 3) in cancerous tissues was higher than the normal colon mucosa from the same patient (p < 0.05). There was no difference in the MyD88 and IFN-α gene expression between cancerous and normal colon mucosa.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Division of Colorectal Surgery, Department of Surgery, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung 833, Taiwan. cclu999@gmail.com.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus