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Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy.

Furukawa J, Tsuda M, Okada K, Kimura T, Piao J, Tanaka S, Shinohara Y - PLoS ONE (2015)

Bottom Line: Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model.More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified.The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT).

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, and Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, Sapporo, Japan.

ABSTRACT
Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response.

No MeSH data available.


Related in: MedlinePlus

Model NHA cells and experimental design for the glycomic analysis.(a) Generation of immortalized and transformed human astrocytes. (b) Expression levels of specific defined elements (i.e., hTERT, SV40ER (both large T and small t), H-RasV12, and myrAKT) assessed by immunoblotting. (c) Protocols employed for glycomic analyses of N-, O- and GSL-glycans. For further details, see Methods.
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pone.0128300.g001: Model NHA cells and experimental design for the glycomic analysis.(a) Generation of immortalized and transformed human astrocytes. (b) Expression levels of specific defined elements (i.e., hTERT, SV40ER (both large T and small t), H-RasV12, and myrAKT) assessed by immunoblotting. (c) Protocols employed for glycomic analyses of N-, O- and GSL-glycans. For further details, see Methods.

Mentions: We employed our original brain tumor model in which parental NHAs were transformed through the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, and designated the transformed NHAs as NHA/T, NHA/TS, NHA/TSR, and NHA/TSRA cells, respectively. Although the NHA/TS cells exhibited a higher rate of cell growth than the parental NHAs, they failed to form colonies in soft agar, indicating their status as immortalized but not transformed. In our previous study, we found that NHA/TSR cells successfully formed a considerable number of colonies in soft agar, along with xenografted tumors in nude mice [20]. Intratumoral necrosis was displayed in the context of xenografted tumors formed by NHA/TSRA cells, with a higher proliferative index. Thus, NHA/TS cells appear to mimic benign grade II gliomas, whereas NHA/TSR and NHA/TSRA cells exhibit histopathological features of malignant grade III and IV gliomas, respectively [20] (Fig 1a).


Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy.

Furukawa J, Tsuda M, Okada K, Kimura T, Piao J, Tanaka S, Shinohara Y - PLoS ONE (2015)

Model NHA cells and experimental design for the glycomic analysis.(a) Generation of immortalized and transformed human astrocytes. (b) Expression levels of specific defined elements (i.e., hTERT, SV40ER (both large T and small t), H-RasV12, and myrAKT) assessed by immunoblotting. (c) Protocols employed for glycomic analyses of N-, O- and GSL-glycans. For further details, see Methods.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488535&req=5

pone.0128300.g001: Model NHA cells and experimental design for the glycomic analysis.(a) Generation of immortalized and transformed human astrocytes. (b) Expression levels of specific defined elements (i.e., hTERT, SV40ER (both large T and small t), H-RasV12, and myrAKT) assessed by immunoblotting. (c) Protocols employed for glycomic analyses of N-, O- and GSL-glycans. For further details, see Methods.
Mentions: We employed our original brain tumor model in which parental NHAs were transformed through the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, and designated the transformed NHAs as NHA/T, NHA/TS, NHA/TSR, and NHA/TSRA cells, respectively. Although the NHA/TS cells exhibited a higher rate of cell growth than the parental NHAs, they failed to form colonies in soft agar, indicating their status as immortalized but not transformed. In our previous study, we found that NHA/TSR cells successfully formed a considerable number of colonies in soft agar, along with xenografted tumors in nude mice [20]. Intratumoral necrosis was displayed in the context of xenografted tumors formed by NHA/TSRA cells, with a higher proliferative index. Thus, NHA/TS cells appear to mimic benign grade II gliomas, whereas NHA/TSR and NHA/TSRA cells exhibit histopathological features of malignant grade III and IV gliomas, respectively [20] (Fig 1a).

Bottom Line: Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model.More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified.The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT).

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, and Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, Sapporo, Japan.

ABSTRACT
Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response.

No MeSH data available.


Related in: MedlinePlus