-1,3-Galactosyl-O-Glycosyl-Glycoprotein -1,6-N-Acetylglucosaminyltransferase 3 Increases MCAM Stability, Which Enhances S100A8/A9-Mediated Cancer Motility

Abstract

We previously identified novel S100A8/A9 receptors, extracellular matrix metalloproteinase inducer (EMMPRIN), melanoma cell adhesion molecule (MCAM), activated leukocyte cell adhesion molecule (ALCAM), and neuroplastin (NPTN) β, that are critically involved in S100A8/A9-mediated cancer metastasis and inflammation when expressed at high levels. However, little is known about the presence of any cancer-specific mechanism(s) that modifies these receptors, further inducing upregulation at protein levels without any transcriptional regulation. Expression levels of glycosyltransferase-encoding genes were examined by a PCR-based profiling array followed by confirmation with quantitative real-time PCR. Cell migration and invasion were assessed using a Boyden chamber. Western blotting was used to examine the protein level, and the RNA level was examined by Northern blotting. Immunohistochemistry was used to examine the expression pattern of β-1,3-galactosyl--glycosyl-glycoprotein β-1,6--acetylglucosaminyltransferase 3 (GCNT3) and MCAM in melanoma tissue. We found that GCNT3 is overexpressed in highly metastatic melanomas. Silencing and functional inhibition of GCNT3 greatly suppressed migration and invasion of melanoma cells, resulting in the loss of S100A8/A9 responsiveness. Among the novel S100A8/A9 receptors, GCNT3 favorably glycosylates the MCAM receptor, extending its half-life and leading to further elevation of S100A8/A9-mediated cellular motility in melanoma cells. GCNT3 expression is positively correlated to MCAM expression in patients with high-grade melanomas. Collectively, our results showed that GCNT3 is an upstream regulator of MCAM protein and indicate the possibility of a potential molecular target in melanoma therapeutics through abrogation of the S100A8/A9-MCAM axis.

Neuroplastin‐β mediates S100A8/A9‐induced lung cancer disseminative progression

“…We also established several A549 stable clones that showed indefinitely stable expression of the aberrant genes of interest at much higher levels. Based on the pIDT‐SMART (C‐TSC) vector, we made two improved vectors for stable expression with significantly higher level, named pSAKA‐1B and pSAKA‐4B . The expression vector pSAKA‐1B is specifically adapted to Chinese hamster ovary (CHO) cells, while pSAKA‐4B is useful for a wide range of diverse cell lines from different kinds of mammalian species in a constant manner.…”

“…Besides classical S100A8/A9 receptors, toll-like receptor 4 (TLR4) 4,5 and receptor for advanced glycation end products (RAGE), 5,6 we have reported the presence of the important novel receptors including extracellular matrix metalloproteinase inducer (EMMPRIN), neuroplastin (NPTN)α and β, melanoma cell adhesion molecule (MCAM) and activated leukocyte cell adhesion molecule (ALCAM). [7][8][9][10][11][12] Among these receptors, we found that lung cancer cell lines highly express NPTNβ compared with normal cells lines ( Figure S1A). This finding was supported by the result of immunohistochemistry showing that NPTNβ was highly positive in lung cancer cells and that S100A8/A9 appeared in either cancer cells or the surrounding stroma in clinical specimens, suggesting that extracellular S100A8/A9 physiologically acts on cell surface NPTNβ in cancer cells in an autocrine manner as well as a paracrine manner ( Figure 1).…”

Newly developed anti‐S100A8/A9 monoclonal antibody efficiently prevents lung tropic cancer metastasis

“…Namely, extracellular S100A8/A9 secreted from the lungs attracts melanoma cells from distant skin foci, acting as strong ‘soil signals’ to receptors in the cancer cells that function as ‘S100A8/A9‐soil signal sensors’. These sensors include the renowned Toll‐like receptor 4 (TLR4), receptor for advanced glycation end products (RAGE), and novel receptors that we have identified, MCAM, ALCAM, EMMPRIN and NPTNα and β.…”

exSSSRs (extracellular S100 soil sensor receptors)‐Fc fusion proteins work as prominent decoys to S100A8/A9‐induced lung tropic cancer metastasis

“…2 We then found further evidence of other important novel soil sensors for S100A8/A9: EMMPRIN, 3 NPTNβ, 4 MCAM and ALCAM. 5,6 We named the proteins "S100 soil sensor receptors, abbreviated as SSSRs." 5,6 The relationship between S100A8/A9 and these receptors may be applicable to broad cancer species since S100A8/A9 is also utilized by breast cancer cells for lung tropic metastasis.…”

“…5,6 We named the proteins "S100 soil sensor receptors, abbreviated as SSSRs." 5,6 The relationship between S100A8/A9 and these receptors may be applicable to broad cancer species since S100A8/A9 is also utilized by breast cancer cells for lung tropic metastasis. 7 Since these receptors seem to be differently expressed depending on the cancer species and since their expression seems to be upregulated in response to the progression of cancer malignancy (our study and our ongoing studies), the S100A8/A9-SSSRs axis therefore may act as an important mediator in "seed and soil" metastasis in multiple cancer species.…”