Evaluation of 2-Thioxoimadazolidin-4-one Derivatives as Potent Anti-Cancer Agents through Apoptosis Induction and Antioxidant Activation: In Vitro and In Vivo Approaches

• Published in: Molecules (Basel, Switzerland) Switzerland), 2021-12, Vol.27 (1), p.83
• Main Authors: Nafie, Mohamed S, Khodair, Ahmed I, Hassan, Hebat Allah Y, El-Fadeal, Noha M Abd, Bogari, Hanin A, Elhady, Sameh S, Ahmed, Safwat A
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; 2-thioxoimadazolidin-4-one; AKT protein; Animals; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacology; antioxidant activity; Antioxidants; Antioxidants - chemistry; Antioxidants - pharmacology; Antitumor activity; Apoptosis; Apoptosis - drug effects; Bcl-2 protein; Cancer therapies; Caspase-3; Cell cycle; Cell division; Cell growth; Cell Line, Tumor; Cell Proliferation - drug effects; Cytotoxicity; Disease Models, Animal; Flow cytometry; Gene expression; Gene Expression Regulation, Neoplastic - drug effects; Hematology; Hep G2 Cells; Hepatocellular carcinoma; Hepatocytes; Humans; Imidazolidines - chemistry; Imidazolidines - pharmacology; in vivo; In vivo methods and tests; Investigations; Kinases; Liver cancer; Liver diseases; Mice; Models, Molecular; Molecular Conformation; Molecular docking; Molecular Structure; p53 Protein; Phosphatidylinositol 3-Kinases - metabolism; PI3K/AKT; Polymerase chain reaction; Protein Binding; Proteins; Proto-Oncogene Proteins c-akt - metabolism; Signal Transduction; Staurosporine; Structure-Activity Relationship; Western blotting; Xenograft Model Antitumor Assays
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules27010083

Hepatocellular carcinoma (HCC) is one of the most widespread malignancies and is reported as the fourth most prevalent cause of cancer deaths worldwide. Therefore, we aimed to investigate the probable mechanistic cytotoxic effect of the promising 2-thioxoimidazolidin-4-one derivative on liver cancer cells using in vitro and in vivo approaches. The compounds were tested for the in vitro cytotoxic activity using MTT assay, and the promising compound was tested in colony forming unit assay, flow cytometric analysis, RT-PCR, Western blotting, in vivo using SEC-carcinoma and in silico to highlight the virtual mechanism of action. Both compounds and performed cytotoxic effects against HepG2 cells with IC values of 0.017 and 0.18 μM, respectively, compared to Staurosporine and 5-Fu as reference drugs with IC values of 5.07 and 5.18 µM, respectively. Compound treatment revealed apoptosis induction by 19.35-fold (11.42% compared to 0.59% in control), arresting the cell cycle at G2/M phase. Moreover, studying gene expression that plays critical roles in cell cycle and apoptosis by RT-PCR demonstrated that compound enhances the expression of the pro-apoptotic genes p53, PUMA, and Caspase 3, 8, and 9, and impedes the anti-apoptotic Bcl-2 gene in the HepG2 cells. It can also inhibit the PI3K/AKT pathway at both gene and protein levels, which was reinforced by the in silico predictions of the molecular docking simulations towards the PI3K/AKT proteins. Finally, in vivo study verified that compound has a promising anti-cancer activity through activating antioxidant levels (CAT, SOD and GSH) and ameliorating hematological, biochemical, and histopathological findings.