3,3’,5,5’-Tetramethoxybiphenyl-4,4’-diol exerts a cytotoxic effect on hepatocellular carcinoma cell lines by inducing morphological and ultrastructural alterations, G2/M cell cycle arrest and death by apoptosis via CDK1 interaction
Hepatocellular carcinoma (HCC) is a highly aggressive malignancy with frequent recurrence and chemoresistance, underscoring the need for new treatment strategies. 3,3',5,5'-Tetramethoxybiphenyl-4,4'-diol (TMBP) showed cytotoxicity against lung cancer cell lines without harming normal cells. Thus, we investigated the antitumoral effect of TMBP on HCC cell lines, HuH7.5 (p53-mutant) and HepG2/C3A (p53-wild type). Cells were treated with TMBP (12.5-150 µM) for 24 and 48 h, and metabolic cellular activity (MTT) were used to determine the 50 % inhibitory concentration (IC<sub>50</sub>) values. TMBP cytotoxicity were assessed by Trypan blue assay, scanning and transmission electron microscopy. Cell migration (wound healing), total ROS (H<sub>2</sub>DCFDA), mitochondrial dysfunction (TMRE), lipid droplets (Nile Red), and autophagic vacuoles (MDC) were assessed. Flow cytometry characterized cell cycle distribution and cell death. Caspase 3/7 activity and CASP3 expression confirmed apoptosis. Molecular docking and gene expression analysis validated TMBP-CDK1 interaction. TMBP reduced cell viability, with IC<sub>50</sub> values of 68 and 55 µM (HuH7.5) and 50 and 42 µM (HepG2/C3A) at 24 and 48 h. TMBP induced severe morphological alterations, impaired migration, increased ROS, mitochondrial dysfunction, increased lipid droplets and autophagic vacuoles. TMBP also led to G2/M arrest and apoptosis, likely via CDK1 inhibition through hydrogen bonding at Tyr15. These findings highlight TMBP as a promising therapeutic candidate targeting CDK1 in HCC.
Authors
- Mariana Barbosa Detoni (ORCID: https://orcid.org/0000-0001-9526-1948)
- Carolina Panis (ORCID: https://orcid.org/0000-0002-0104-4369)
- Danielle Lazarin‐Bidóia (ORCID: https://orcid.org/0000-0003-2579-0048)
- Marcelle de Lima Ferreira Bispo (ORCID: https://orcid.org/0000-0002-6001-360X)
- Kaio Maciel de Santiago-Silva (ORCID: https://orcid.org/0000-0001-6550-7820)
- Jéseka G. Schirmann (ORCID: https://orcid.org/0000-0002-1690-5889)
- Fabricio Seidy Ribeiro Inoue
- Marcelo Assis (ORCID: https://orcid.org/0000-0003-0355-5565)
- Virgínia Márcia Concato (ORCID: https://orcid.org/0000-0001-5451-6254)
- Rayanne Regina Beltrame Machado (ORCID: https://orcid.org/0000-0001-5918-9758)
- Bruna Taciane da Silva Bortoleti (ORCID: https://orcid.org/0000-0003-2631-6719)
- Ellen Mayara Souza Cruz (ORCID: https://orcid.org/0000-0003-2445-5598)
- Robert F. H. Dekker (ORCID: https://orcid.org/0000-0002-3787-6917)
- Mário Sérgio Mantovani (ORCID: https://orcid.org/0000-0001-5268-6508)
- Ivete Conchon‐Costa (ORCID: https://orcid.org/0000-0002-9800-2068)
- Aneli M. Barbosa‐Dekker (ORCID: https://orcid.org/0000-0002-2339-8985)
- Fernanda Tomiotto‐Pellissier (ORCID: https://orcid.org/0000-0002-7273-8237)
- Manoela Daiele Gonçalves-Lens
- Wander Rogério Pavanelli (ORCID: https://orcid.org/0000-0003-1734-7038)
- Juliana Maria Bitencourt de Morais-Valentim
Institutions
- Universidade Estadual de Maringá (BR)
- Universidade Estadual de Londrina (BR)
- Universidade Tecnológica Federal do Paraná (BR)
- Universidade do Estado do Pará (BR)
Publication Details
- Journal
- Biomedicine & Pharmacotherapy
- Published
- 2025-04-24
- DOI
- https://doi.org/10.1016/j.biopha.2025.118082
- Citations
- 1
- Primary Topic
- Cancer Mechanisms and Therapy
- Type
- article
- Field-Weighted Citation Impact
- 1.47
Funders
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
- Conselho Nacional de Desenvolvimento Científico e Tecnológico
- Financiadora de Estudos e Projetos
- Universidade Estadual de Londrina