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Int. J. Mol. Sci. 2016, 17(5), 652; doi:10.3390/ijms17050652

Exogenous and Endogeneous Disialosyl Ganglioside GD1b Induces Apoptosis of MCF-7 Human Breast Cancer Cells

1
Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea
2
Research Institute, Davinch-K Co., Ltd., B1603-3, 606, Seobusaet-gil, Geumcheon-gu, Seoul 153-719, Korea
3
Division of Enteric Diseases, Center for Infectious Diseases Research, Korea National Institute of Health, Heungdeok-gu, Cheongju 363-951, Korea
4
Functional Genomics Laboratory, Department of Animal Sciences, the Ohio State University, 2029 Fyffe Court, Columbus, OH 43210, USA
5
Research Institute of Biomedical Engineering and Department of Medicine, Catholic University of Daegu School of Medicine, Daegu 705-718, Korea
6
Faculty of Medicinal Biotechnology, Dong-A University, Busan 604-714, Korea
7
Division of Applied Medicine, School of Korean Medicine, Pusan National University, Yangsan City, Gyeongsangnam-Do 626-870, Korea
8
College of Pharmacy, Yeungnam University, Gyeongsan 712-749, Korea
9
Department of Medical Device Management and Research, Samsung Advanced Institute for Health Sciences & Technology (SAIHST), Seoul 06351, Korea
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Ester Boix
Received: 22 March 2016 / Revised: 13 April 2016 / Accepted: 21 April 2016 / Published: 30 April 2016
(This article belongs to the Special Issue Glycan–Receptor Interaction)
View Full-Text   |   Download PDF [3184 KB, uploaded 30 April 2016]   |  

Abstract

Gangliosides have been known to play a role in the regulation of apoptosis in cancer cells. This study has employed disialyl-ganglioside GD1b to apoptosis in human breast cancer MCF-7 cells using exogenous treatment of the cells with GD1b and endogenous expression of GD1b in MCF-7 cells. First, apoptosis in MCF-7 cells was observed after treatment of GD1b. Treatment of MCF-7 cells with GD1b reduced cell growth rates in a dose and time dependent manner during GD1b treatment, as determined by XTT assay. Among the various gangliosides, GD1b specifically induced apoptosis of the MCF-7 cells. Flow cytometry and immunofluorescence assays showed that GD1b specifically induces apoptosis in the MCF-7 cells with Annexin V binding for apoptotic actions in early stage and propidium iodide (PI) staining the nucleus of the MCF-7 cells. Treatment of MCF-7 cells with GD1b activated apoptotic molecules such as processed forms of caspase-8, -7 and PARP (Poly(ADP-ribose) polymerase), without any change in the expression of mitochondria-mediated apoptosis molecules such as Bax and Bcl-2. Second, to investigate the effect of endogenously produced GD1b on the regulation of cell function, UDP-gal: β1,3-galactosyltransferase-2 (GD1b synthase, Gal-T2) gene has been transfected into the MCF-7 cells. Using the GD1b synthase-transfectants, apoptosis-related signal proteins linked to phenotype changes were examined. Similar to the exogenous GD1b treatment, the cell growth of the GD1b synthase gene-transfectants was significantly suppressed compared with the vector-transfectant cell lines and transfection activated the apoptotic molecules such as processed forms of caspase-8, -7 and PARP, but not the levels of expression of Bax and Bcl-2. GD1b-induced apoptosis was blocked by caspase inhibitor, Z-VAD. Therefore, taken together, it was concluded that GD1b could play an important role in the regulation of breast cancer apoptosis. View Full-Text
Keywords: disialyl-ganglioside GD1b; human breast cancer MCF-7 cells; apoptosis; caspase; human β1,3-galactosyltransferase-2 (GD1b synthase; Gal-T2) disialyl-ganglioside GD1b; human breast cancer MCF-7 cells; apoptosis; caspase; human β1,3-galactosyltransferase-2 (GD1b synthase; Gal-T2)
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Ha, S.-H.; Lee, J.-M.; Kwon, K.-M.; Kwak, C.-H.; Abekura, F.; Park, J.-Y.; Cho, S.-H.; Lee, K.; Chang, Y.-C.; Lee, Y.-C.; Choi, H.-J.; Chung, T.-W.; Ha, K.-T.; Chang, H.-W.; Kim, C.-H. Exogenous and Endogeneous Disialosyl Ganglioside GD1b Induces Apoptosis of MCF-7 Human Breast Cancer Cells. Int. J. Mol. Sci. 2016, 17, 652.

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