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. 2020 Feb 9;9(2):397. doi: 10.3390/cells9020397

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Identification of BGN as a potential TLR4 ligand involved in suppression of Siglec ligands in the early stage of colon cancer carcinogenesis. (A) Gene expression analysis of TLR4 ligands—BGN, HSPD1, HMGB1, S100A8, S100A9, TNC, heparan sulfate proteoglycan active enzymes (SDC2 or HSPG2), hyaluronidases (HYAL1 and HYAL2) and hyaluronan synthase (HAS1, HAS2, and HAS3)—in normal (n = 397) and colon cancer samples (n = 3775) from the GENT2 database. ***, p < 0.001. Results are shown as the relative fold increase compared with normal colon control. (B) Comparison of BGN, S100A8, S100A9, and TNC amount expression in COAD and READ by GEPIA2. Gene expression level was calculated by transcript per million (TPM). (C) Large-scale RNA-Seq transcriptome analysis of BGN in healthy colorectal tissues and tissues from four stages of colorectal cancer from the TCGA database. n, number of patients; *** p < 0.001. (D) BGN was inversely correlated with the overall survival of patients with colorectal cancer. Overall survival analyses were performed using the GEPIA2 online platform. The solid line shows the survival curve, and the dotted line shows the 95% confidence interval. Red lines indicate patients with expression above the median, and blue lines indicate patients with expression below the median. Log-rank p < 0.05 was considered statistically significant. HR, hazard ratio. (E) BGN expression in HT-29 and LS174T after the administration of shRNAs for BGN. (F) Flow cytometry analysis of expression of disialyl Lewis^a on HT-29 cells (top panels) and sialyl 6- sulfo Lewis^x on LS174T cells (bottom panels), after the administration of shRNAs for BGN. (G) Fluorescence intensity results of flow cytometry analysis in (H) are shown. The relative expression levels of disialyl Lewis^a and sialyl 6-sulfo Lewis^x are shown as mean fluorescence intensity (MFI) compared with the staining of control IgG or IgM. *** p < 0.001. (H) Restoration of Siglec-7 binding by introduction of shRNAs for BGN in LS174T colon cancer cells. The relative levels of Siglec-7 are shown as the relative mean fluorescence intensity (MFI) compared with the control group IgG. *** p < 0.001.

Identification of BGN as a potential TLR4 ligand involved in suppression of Siglec ligands in the early stage of colon cancer carcinogenesis. (A) Gene expression analysis of TLR4 ligands—BGN, HSPD1, HMGB1, S100A8, S100A9, TNC, heparan sulfate proteoglycan active enzymes (SDC2 or HSPG2), hyaluronidases (HYAL1 and HYAL2) and hyaluronan synthase (HAS1, HAS2, and HAS3)—in normal (n = 397) and colon cancer samples (n = 3775) from the GENT2 database. ***, p < 0.001. Results are shown as the relative fold increase compared with normal colon control. (B) Comparison of BGN, S100A8, S100A9, and TNC amount expression in COAD and READ by GEPIA2. Gene expression level was calculated by transcript per million (TPM). (C) Large-scale RNA-Seq transcriptome analysis of BGN in healthy colorectal tissues and tissues from four stages of colorectal cancer from the TCGA database. n, number of patients; *** p < 0.001. (D) BGN was inversely correlated with the overall survival of patients with colorectal cancer. Overall survival analyses were performed using the GEPIA2 online platform. The solid line shows the survival curve, and the dotted line shows the 95% confidence interval. Red lines indicate patients with expression above the median, and blue lines indicate patients with expression below the median. Log-rank p < 0.05 was considered statistically significant. HR, hazard ratio. (E) BGN expression in HT-29 and LS174T after the administration of shRNAs for BGN. (F) Flow cytometry analysis of expression of disialyl Lewis^a on HT-29 cells (top panels) and sialyl 6- sulfo Lewis^x on LS174T cells (bottom panels), after the administration of shRNAs for BGN. (G) Fluorescence intensity results of flow cytometry analysis in (H) are shown. The relative expression levels of disialyl Lewis^a and sialyl 6-sulfo Lewis^x are shown as mean fluorescence intensity (MFI) compared with the staining of control IgG or IgM. *** p < 0.001. (H) Restoration of Siglec-7 binding by introduction of shRNAs for BGN in LS174T colon cancer cells. The relative levels of Siglec-7 are shown as the relative mean fluorescence intensity (MFI) compared with the control group IgG. *** p < 0.001.