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Catfish rhamnose-binding lectin induces G0/1 cell cycle arrest in Burkitt’s lymphoma cells via membrane surface Gb3

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Abstract

Silurus asotus egg lectin (SAL), an α-galactoside-binding protein isolated from the eggs of catfish, is a member of the rhamnose-binding lectin family that binds to Gb3 glycan (Galα1–4Galβ1–4Glc). We have previously demonstrated that SAL reduces the proliferation of Gb3-expressing Burkitt’s lymphoma Raji cells and confirm here that it does not reduce their viability, indicating that unlike other lectins, it is not cytotoxic. The aim of this study was to determine the signal transduction mechanism(s) underlying this novel SAL/Gb3 binding-mediated effect profile. SAL/Gb3 interaction arrested the cell cycle through increasing the G0/1 phase population of Raji cells. SAL suppressed the transcription of cell cycle-related factors such as c-MYC, cyclin D3, and cyclin-dependent protein kinase (CDK)-4. Conversely, the CDK inhibitors p21 and p27 were elevated by treatment with SAL. In particular, the production of p27 in response to SAL treatment increased steadily, whereas p21 production was maximal at 12 h and lower at 24 h. Activation of Ras-MEK-ERK pathway led to an increase in expression of p21. Notably, treatment of Raji cells with anti-Gb3 mAb alone did not produce the above effects. Taken together, our findings suggest that Gb3 on the Raji cell surface interacts with SAL to trigger sequential GDP-Ras phosphorylation, Ras-MEK-ERK pathway activation, p21 production, and cell cycle arrest at the G0/1 phase.

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Abbreviations

BrdU:

Bromodeoxy uridine

CCND3:

Cyclin D3

CDK:

Cyclin-dependent protein kinase

ERK:

Extracellular signal-regulated kinase

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

Gb3:

Galα1–4Galβ1–4Glc-Cer

GDP:

Guanosine diphosphate

GEM:

Glycosphingolipid-enriched microdomains

GTP:

Guanosine triphosphate

GSL:

Glycosphingolipid

mAb:

Monoclonal antibody

JNK:

c-Jun N-terminal kinase

MAPK:

Mitogen-activated protein kinase

MEK:

MAPK/ERK kinase

PVDF:

Polyvinylidene difluoride

qRT-PCR:

Quantitative reverse transcription-polymerase chain reaction

SAL:

Silurus asotus egg lectin

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SUEL:

Sea urchin egg lectin

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Acknowledgments

This study was supported by the “Academic Frontier” Project for Private Universities and the “Strategic Project to Support the Formation of Research Bases at Private Universities (SENRYAKU)” from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT), Japan. We would like to thank Editage (www.editage.jp) for English language editing.

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Authors and Affiliations

  1. Division of Cell Recognition Study, Institute of Molecular Biomembrane and Glycobiology, Tohoku Medical and Pharmaceutical University, 4-4-1 Komatsushima, Aoba-ku, Sendai, 981-8558, Japan

    Shigeki Sugawara, Changhun Im, Tasuku Kawano, Takeo Tatsuta, Kazuo Nitta & Masahiro Hosono

  2. Laboratory of Glycobiology and Marine Biochemistry, Department of Life and Environmental System Science, Graduate School of NanoBio Sciences, Yokohama City University, 22-2 Seto, Kanazawa-ku, Yokohama, 236-0027, Japan

    Yasuhiro Koide, Daiki Yamamoto & Yasuhiro Ozeki

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  1. Shigeki Sugawara
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  2. Changhun Im
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  3. Tasuku Kawano
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  4. Takeo Tatsuta
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  5. Yasuhiro Koide
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  7. Yasuhiro Ozeki
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  8. Kazuo Nitta
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  9. Masahiro Hosono
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Correspondence to Masahiro Hosono.

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Sugawara, S., Im, C., Kawano, T. et al. Catfish rhamnose-binding lectin induces G0/1 cell cycle arrest in Burkitt’s lymphoma cells via membrane surface Gb3. Glycoconj J 34, 127–138 (2017). https://doi.org/10.1007/s10719-016-9739-2

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