Overexpression and β-1,6-N-Acetylglucosaminylation-initiated Aberrant Glycosylation of TIMP-1

Kim, Yong-Sam; Ahn, Yeong Hee; Song, Kyoung; Kang, Jeong Gu; Lee, Ju Hee; Jeon, Seong Kook; Kim, Hyoung-Chin; Yoo, Jong Shin; Ko, Jeong–Heon

doi:10.1074/jbc.m112.370064

Cited by 35 publications

(26 citation statements)

References 49 publications

(34 reference statements)

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“…Despite the increase in Timp1 expression in 4C11- and 4C11+ melanoma cell lines, high MMP activity was found in these melanoma cell lines (Figure 1D), suggesting MMP-independent functions of Timp1. Recently, Kim and coworkers showed that colon cancer cells have TIMP1 molecule bearing aberrant glycosylation, which impairs its efficient function as MMP inhibitor [33]. In fact, an increase in N- linked oligosaccharides was observed in melanoma cells derived from melan-a subjected to sequential cycles of deadhesion [10], reinforcing the idea that TIMP1 would confer tumor aggressiveness independent of its function on MMP activity.…”

Section: Discussionmentioning

confidence: 99%

Timp1 interacts with beta-1 integrin and CD63 along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway independently of Akt phosphorylation

et al. 2013

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BackgroundAnoikis resistance is one of the abilities acquired along tumor progression. This characteristic is associated with metastasis development, since tumorigenic cells must survive independently of cell-matrix interactions in this process. In our laboratory, it was developed a murine melanocyte malignant transformation model associated with a sustained stressful condition. After subjecting melan-a melanocytes to 1, 2, 3 and 4 cycles of anchorage impediment, anoikis resistant cells were established and named 1C, 2C, 3C and 4C, respectively. These cells showed altered morphology and PMA independent cell growth, but were not tumorigenic, corresponding to pre-malignant cells. After limiting dilution of 4C pre-malignant cells, melanoma cell lines with different characteristics were obtained. Previous data from our group showed that increased Timp1 expression correlated with anoikis-resistant phenotype. Timp1 was shown to confer anchorage-independent growth capability to melan-a melanocytes and render melanoma cells more aggressive when injected into mice. However, the mechanisms involved in anoikis regulation by Timp1 in tumorigenic cells are not clear yet.MethodsThe β1-integrin and Timp1 expression were evaluated by Western blotting and CD63 protein expression by flow cytometry using specific antibodies. To analyze the interaction among Timp1, CD63 and β1-integrin, immunoprecipitation assays were performed, anoikis resistance capability was evaluated in the presence or not of the PI3-K inhibitors, Wortmannin and LY294002. Relative expression of TIMP1 and CD63 in human metastatic melanoma cells was analyzed by real time PCR.ResultsDifferential association among Timp1, CD63 and β1-integrins was observed in melan-a melanocytes, 4C pre-malignant melanocytes and 4C11- and 4C11+ melanoma cells. Timp1 present in conditioned medium of melanoma cells rendered melan-a melanocytes anoikis-resistant through PI3-K signaling pathway independently of Akt activation. In human melanoma cell lines, in which TIMP1 and beta-1 integrin were also found to be interacting, TIMP1 and CD63 levels together was shown to correlate significantly with colony formation capacity.ConclusionsOur results show that Timp1 is assembled in a supramolecular complex containing CD63 and β1-integrins along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway, independently of Akt phosphorylation. In addition, our data point TIMP1, mainly together with CD63, as a potential biomarker of melanoma.

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Section: Discussionmentioning

confidence: 99%

Timp1 interacts with beta-1 integrin and CD63 along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway independently of Akt phosphorylation

et al. 2013

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“…These proteins are all heavily glycosylated proteins and known to be involved in resistance to chemotherapies in multiple cancer types [ 47 – 52 ], [ 16 , 53 , 54 ]. Moreover, as glycoproteins, the functions of TIMP1 and CLU were reported to be markedly influenced by altered glycans [ 55 – 57 ]. Nevertheless, previous studies rarely focused on the alterations of protein glycosylation during multi-drug resistance in cancer.…”

Section: Discussionmentioning

confidence: 99%

Characterization of site-specific glycosylation of secreted proteins associated with multi-drug resistance of gastric cancer

Qin

et al. 2016

Oncotarget

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Multi-drug resistance (MDR) remains a great obstacle to effective chemotherapy for gastric cancer. A number of secreted glycoproteins have been reported to be involved in the development of MDR in gastric cancer. However, whether glycosylation of secreted glycoproteins changes during MDR of gastric cancer is unclear. Our present work manifested that N-glycosites and site-specific glycoforms of secreted proteins in drug-resistant cell lines were distinctly different from those in the parental cell line for the first time. Further characterization highlighted the significance of some aberrantly glycosylated secretory proteins in MDR, suggesting that manipulating the glycosylation of specific glycoproteins could be a potential target for overcoming multi-drug resistance in gastric cancer.

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“…Colon cancer cells are known to exhibit abnormal glycosylation of proteins 34, 35 . We used a glycosylation mutant that showed substantially reduced complex glycan modification and defective cell surface expression to model the impact ofSIGIRR ΔE8 in tumorigenesis.…”

Section: Discussionmentioning

confidence: 99%

Human Colon Tumors Express a Dominant-Negative Form of SIGIRR That Promotes Inflammation and Colitis-Associated Colon Cancer in Mice

et al. 2015

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Background & Aims Single immunoglobulin and toll-interleukin 1 receptor (SIGIRR), a negative regulator of the Toll-like and interleukin-1 receptor (IL1R) signaling pathways, controls intestinal inflammation and suppresses colon tumorigenesis in mice. However, the importance of SIGIRR in human colorectal cancer development has not been determined. We investigated the role of SIGIRR in development of human colorectal cancer. Methods We performed RNA sequence analyses of pairs of colon tumor and non-tumor tissues, each collected from 68 patients. Immunoblot and immunofluorescence analyses were used to determine levels of SIGIRR protein in primary human colonic epithelial cells, tumor tissues, and colon cancer cell lines. We expressed SIGIRR and mutant forms of the protein in Vaco cell lines. We created and analyzed mice that expressed full-length (control) or a mutant form of Sigirr (encoding SIGIRRN86/102S, which is not glycosylated) specifically in the intestinal epithelium. Some mice were given azoxymethane and dextran sulfate sodium to induce colitis-associated cancer. Intestinal tissues were collected and analyzed by immunohistochemical and gene expression profile analyses. Results RNA sequence analyses revealed increased expression of a SIGIRR mRNA isoform, SIGIRRΔE8, in colorectal cancer tissues compared to paired non-tumor tissues. SIGIRRΔE8 is not modified by complex glycans and is therefore retained in the cytoplasm—it cannot localize to the cell membrane or reduce IL1R signaling. SIGIRRΔE8 interacts with and has a dominant-negative effect on SIGIRR, reducing its glycosylation, localization to the cell surface, and function. Most SIGIRR detected in human colon cancer tissues was cytoplasmic, whereas in non-tumor tissues it was found at the cell membrane. Mice that expressed SIGIRRN86/102S developed more inflammation and formed larger tumors after administration of azoxymethane and dextran sulfate sodium than control mice; colon tissues from these mutant mice expressed higher levels of the inflammatory cytokines IL17A and IL6 had activation of the transcription factors STAT3 and NFκB. SIGIRRN86/102S expressed in colons of mice did not localize to the epithelial cell surface. Conclusion Levels of SIGIRR are lower in human colorectal tumors, compared with non-tumor tissues; tumors contain the dominant-negative isoform SIGIRRΔE8. This mutant protein blocks localization of full-length SIGIRR to the surface of colon epithelial cells and its ability to downregulate IL1R signaling. Expression of SIGIRRN86/102S in the colonic epithelium of mice increases expression of inflammatory cytokines and formation and size of colitis-associated tumors.

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Overexpression and β-1,6-N-Acetylglucosaminylation-initiated Aberrant Glycosylation of TIMP-1

Cited by 35 publications

References 49 publications

Timp1 interacts with beta-1 integrin and CD63 along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway independently of Akt phosphorylation

Timp1 interacts with beta-1 integrin and CD63 along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway independently of Akt phosphorylation

Characterization of site-specific glycosylation of secreted proteins associated with multi-drug resistance of gastric cancer

Human Colon Tumors Express a Dominant-Negative Form of SIGIRR That Promotes Inflammation and Colitis-Associated Colon Cancer in Mice

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