title: Tideglusib induces apoptosis in human neuroblastoma IMR32 cells, provoking sub-G0/G1 accumulation and ROS generation.
abstract: Neuroblastoma is the most common tumor amongst children amounting to nearly 15% of cancer deaths. This cancer is peculiar in its characteristics, exhibiting differentiation, maturation and metastatic transformation leading to poor prognosis and low survival rates among children. Chemotherapy, though toxic to normal cells, has shown to improve the survival of the patient with emphasis given more towards targeting angiogenesis. Recently, Tideglusib was designed as an 'Orphan Drug' to target the neurodegenerative Alzheimer's disease and gained significant momentum in its function during clinical trials. Duffy et al. recently reported a reduction in cell viability of human IMR32 neuroblastoma cells when treated with Tideglusib at varying concentrations. We investigated the effects of Tideglusib, at various concentrations, compared to Lithium chloride at various concentrations, on IMR32 cells. Lithium, a known GSK-3 inhibitor, was used as a standard to compare the efficiency of Tideglusib in a dose-dependent manner. Cell viability was assessed by MTT assay. The stages of apoptosis were evaluated by AO/EB staining and nuclear damage was determined by Hoechst 33258 staining. Reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) were assessed by DCFDA dye and Rhodamine-123 dye, respectively. Tideglusib reported a significant dose-dependent increase in pro-apoptotic proteins (PARP, Caspase-9, Caspase-7, Caspase-3) and tumor-related genes (FasL, TNF-α, Cox-2, IL-8, Caspase-3). Anti-GSK3 β, pGSK3 β, Bcl-2, Akt-1, p-Akt1 protein levels were observed with cells exposed to Tideglusib and Lithium chloride. No significant dose-dependent changes were observed for the mRNA expression of collagenase MMP-2, the tumor suppressor p53, or the cell cycle protein p21. Our study also reports Tideglusib reducing colony formation and increasing the level of sub-G0/G1 population in IMR32 cells. Our investigations report the significance of Tideglusib as a promising apoptotic inducer in human neuroblastoma IMR32 cells. Our study also reports that LiCl reduced cell viability in IMR32 cells inducing apoptosis mediated by ROS generation.
authors.affiliation: Department of Cellular and Molecular Biochemistry, Frontier Mediville (A Unit of Frontier Lifeline and Dr. K. M. Cherian Heart Foundation), Affiliated to University of Madras, Chennai 601201, Tamil Nadu, India.
authors.forename: Theodore Lemuel
authors.initials: TL
authors.lastname: Mathuram
authors.affiliation: Department of Biochemistry, School of Life sciences, Bharathidasan University, Tiruchirapalli 620024, Tamil Nadu, India.
authors.forename: Vilwanathan
authors.initials: V
authors.lastname: Ravikumar
authors.affiliation: Sealy Center for Vaccine Development, World Health Organization Collaborating Center for Vaccine Research, Evaluation and Training on Emerging Infectious Diseases, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Reece
authors.affiliation: Department of Biochemistry, School of Life sciences, Bharathidasan University, Tiruchirapalli 620024, Tamil Nadu, India.
authors.forename: Selvaraju
authors.initials: S
authors.lastname: Karthik
authors.affiliation: Department of Cellular and Molecular Biochemistry, Frontier Mediville (A Unit of Frontier Lifeline and Dr. K. M. Cherian Heart Foundation), Affiliated to University of Madras, Chennai 601201, Tamil Nadu, India. Electronic address: sheelsasic@yahoo.co.in.
authors.forename: Changam Sheela
authors.initials: CS
authors.lastname: Sasikumar
authors.affiliation: Department of Cardiothoracic Surgery, Frontier Lifeline Hospital, Chennai 600101, Tamil Nadu, India.
authors.forename: Kotturathu Mammen
authors.initials: KM
authors.lastname: Cherian
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D016193:G1 Phase / Q000187:drug effects*; D006801:Humans; D018021:Lithium Chloride / Q000494:pharmacology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D016192:Resting Phase, Cell Cycle / Q000187:drug effects*; D013830:Thiadiazoles / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D017382:Reactive Oxygen Species; D013830:Thiadiazoles; D018021:Lithium Chloride; C520571:tideglusib
keywords: Apoptosis; GSK-3 inhibition; Lithium; Neuroblastoma; ROS; Tideglusib