title: D-galactose induces necroptotic cell death in neuroblastoma cell lines.
abstract: D-Galactose (D-gal) can induce oxidative stress in non-cancer cells and result in cell damage by disturbing glucose metabolism. However, the effect of D-gal on cancer cells is yet to be explored. In this study, we investigated the toxicity of D-gal to malignant cells specifically neuroblastoma cells. As the results, high concentrations of D-gal had significant toxicity to cancer cells, whereas the same concentrations of glucose had no; the viability loss via D-gal treatment was prominent to malignant cells (Neuro2a, SH-SY5Y, PC-3, and HepG2) comparing to non-malignant cells (NIH3T3 and LO(2)). Differing from the apoptosis induced by H(2) O(2), D-gal damaged cells showed the characters of necrotic cell death, such as trypan blue-tangible and early phase LDH leakage. Further experiments displayed that the toxic effect of D-gal can be alleviated by necroptosis inhibitor Necrostatin (Nec-1) and autophagy inhibitor 3-methyladenine (3-MA) but not by caspase inhibitor z-VAD-fmk. D-Gal treatment can transcriptionally up-regulate the genes relevant to necroptosis (Bmf, Bnip3) and autophagy (Atg5, TIGAR) but not the genes related to apoptosis (Caspase3, Bax, and p53). D-Gal did not activate Caspase-3, but prompted puncta-like GFP-LC3 distribution, an indicator for activated autophagy. The involvement of aldose reductase (AR)-mediated polyol pathway was proved because the inhibitor of AR can attenuate the toxicity of D-gal and D-gal treatment elevates the expression of AR. This study demonstrates for the first time that D-gal can induce non-apoptotic but necroptotic cell death in neuroblastoma cells and provides a new clue for developing the strategy against apoptosis-resistant cancers.
authors.affiliation: Lab for Aging Research, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, #1 Keyuan 4 Road, Gaopeng Avenue, High-tech Zone, Chengdu 610041, People's Republic of China.
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.forename: Yangyan
authors.initials: Y
authors.lastname: He
authors.forename: Ling
authors.initials: L
authors.lastname: Wang
authors.forename: Chunfen
authors.initials: C
authors.lastname: Mo
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Junhong
authors.initials: J
authors.lastname: Li
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Liao
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Tang
authors.forename: Hengyi
authors.initials: H
authors.lastname: Xiao
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002448:Cell Adhesion; D016923:Cell Death / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D017931:DNA Primers; D005434:Flow Cytometry; D005690:Galactose / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D009447:Neuroblastoma / Q000473:pathology*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D005690:Galactose