title: Exogenous and Endogeneous Disialosyl Ganglioside GD1b Induces Apoptosis of MCF-7 Human Breast Cancer Cells.
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.
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea. sunspring5@naver.com.
authors.forename: Sun-Hyung
authors.initials: SH
authors.lastname: Ha
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea. jmlee@daum.net.
authors.forename: Ji-Min
authors.initials: JM
authors.lastname: Lee
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea. muscaria80@naver.com.|Research Institute, Davinch-K Co., Ltd., B1603-3, 606, Seobusaet-gil, Geumcheon-gu, Seoul 153-719, Korea. muscaria80@naver.com.
authors.forename: Kyung-Min
authors.initials: KM
authors.lastname: Kwon
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea. hahaaaa@nate.com.
authors.forename: Choong-Hwan
authors.initials: CH
authors.lastname: Kwak
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea. pokusa6@hotmail.com.
authors.forename: Fukushi
authors.initials: F
authors.lastname: Abekura
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea. wnsdud2057@naver.com.
authors.forename: Jun-Young
authors.initials: JY
authors.lastname: Park
authors.affiliation: Division of Enteric Diseases, Center for Infectious Diseases Research, Korea National Institute of Health, Heungdeok-gu, Cheongju 363-951, Korea. skcho38@hotmail.com.
authors.forename: Seung-Hak
authors.initials: SH
authors.lastname: Cho
authors.affiliation: Functional Genomics Laboratory, Department of Animal Sciences, the Ohio State University, 2029 Fyffe Court, Columbus, OH 43210, USA. lee.2626@osu.edu.
authors.forename: Kichoon
authors.initials: K
authors.lastname: Lee
authors.affiliation: Research Institute of Biomedical Engineering and Department of Medicine, Catholic University of Daegu School of Medicine, Daegu 705-718, Korea. ycchang@cu.ac.kr.
authors.forename: Young-Chae
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Faculty of Medicinal Biotechnology, Dong-A University, Busan 604-714, Korea. yclee@dau.ac.kr.
authors.forename: Young-Choon
authors.initials: YC
authors.lastname: Lee
authors.affiliation: Division of Applied Medicine, School of Korean Medicine, Pusan National University, Yangsan City, Gyeongsangnam-Do 626-870, Korea. pure0917@hanmail.net.
authors.forename: Hee-Jung
authors.initials: HJ
authors.lastname: Choi
authors.affiliation: Division of Applied Medicine, School of Korean Medicine, Pusan National University, Yangsan City, Gyeongsangnam-Do 626-870, Korea. chungtw@hanmail.net.
authors.forename: Tae-Wook
authors.initials: TW
authors.lastname: Chung
authors.affiliation: Division of Applied Medicine, School of Korean Medicine, Pusan National University, Yangsan City, Gyeongsangnam-Do 626-870, Korea. haggis@pnu.ac.kr.
authors.forename: Ki-Tae
authors.initials: KT
authors.lastname: Ha
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan 712-749, Korea. hwchang@yu.ac.kr.
authors.forename: Hyeun-Wook
authors.initials: HW
authors.lastname: Chang
authors.affiliation: Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, 300 Chunchun-Dong, Jangan-Gu, Suwon City, Kyunggi-Do 440-746, Korea. chkimbio@skku.edu.|Department of Medical Device Management and Research, Samsung Advanced Institute for Health Sciences &amp; Technology (SAIHST), Seoul 06351, Korea. chkimbio@skku.edu.
authors.forename: Cheorl-Ho
authors.initials: CH
authors.lastname: Kim
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D053179:Caspase 7 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D061945:Caspase Inhibitors / Q000494:pharmacology; D005260:Female; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism; D005732:Gangliosides / Q000096:biosynthesis / Q000633:toxicity*; D006801:Humans; D061986:MCF-7 Cells; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D061945:Caspase Inhibitors; D005732:Gangliosides; D009842:Oligopeptides; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C517629:benzyloxycarbonyl-valyl-alanyl-aspartic acid; C033851:ganglioside, GD1b; D005700:Galactosyltransferases; C053604:UDP-Gal-beta-galactoside beta1-3-galactosyltransferase; D011065:Poly(ADP-ribose) Polymerases; D053179:Caspase 7; D053181:Caspase 8
keywords: Gal-T2); apoptosis; caspase; disialyl-ganglioside GD1b; human breast cancer MCF-7 cells; human β1,3-galactosyltransferase-2 (GD1b synthase