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Norcantharidin combined with 2-deoxy-d-glucose suppresses the hepatocellular carcinoma cells proliferation and
Ragini Singh1, Shuang Cheng1, Jun Li2
1College of Agronomy, Liaocheng University, Liaocheng, 252059 Shandong China.
Abstract:
Present study aims to investigate the combined effect of anticancer drug, norcantharidin (NCTD) in combination with glycolytic inhibitor, i.e. 2-deoxy-d-glucose (2-DG) in liver cancer (HepG2 and Hepa 1-6) cells. Cell viability of NCTD and 2-DG exposed cells was determined by MTT assay, whereas, colony-forming efficiency and migration rate was determined by clonogenic assay and wound healing assay, respectively. Nuclear DAPI staining and Annexin V FITC-PI staining were used to study the apoptosis induction in cells. Fluorescence microscopy imaging was performed to detect the intracellular reactive oxygen species (ROS) generation and mitochondrial membrane potential by staining with DCFDA and JC-1 dye, respectively. Cell viability assay revealed that NCTD and 2-DG exposure in combination displays more cytotoxic effect than a single drug. Additionally, cells lose their colony formation efficiency, as well as the reduced migration rate ability was also observed upon combined exposure. Increased nuclear condensation and mitochondrial membrane depolarization are considered as key features for apoptosis induction in cancerous cells. Furthermore, oxidative stress produced in cells due to enhanced intracellular ROS generation is also major probability for cellular damage. Thus, from the initial data it can be concluded that further preclinical studies will be needed to prove the efficacy of NCTD and 2-DG in hepatocellular carcinoma therapy.
Insights
Combining norcantharidin (NCTD) with 2-deoxy-d-glucose (2-DG) enhances liver cancer cell death. This combination therapy shows greater cytotoxicity and reduced cell migration, indicating potential for hepatocellular carcinoma treatment.
Area of Science:
- Oncology
- Biochemistry
- Cell Biology
Background:
- Hepatocellular carcinoma (HCC) remains a significant global health challenge.
- Targeting cancer cell metabolism, specifically glycolysis, is a promising therapeutic strategy.
- Norcantharidin (NCTD) and 2-deoxy-d-glucose (2-DG) are potential agents for HCC treatment.
Purpose of the Study:
- To investigate the synergistic effects of NCTD and 2-DG on liver cancer cells.
- To evaluate the impact of this combination on cell viability, colony formation, and migration.
- To explore the underlying mechanisms of cell death induction, including apoptosis and oxidative stress.
Main Methods:
- Cell viability was assessed using MTT assays.
- Colony-forming efficiency and migration were evaluated via clonogenic and wound healing assays.
- Apoptosis was detected using DAPI and Annexin V FITC-PI staining.
- Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential were measured using fluorescence microscopy.
Main Results:
- Combined NCTD and 2-DG treatment exhibited a significantly higher cytotoxic effect compared to individual agents.
- The combination therapy led to reduced colony formation efficiency and impaired cell migration.
- Enhanced apoptosis, characterized by nuclear condensation and mitochondrial depolarization, was observed.
- Increased intracellular ROS generation indicated elevated oxidative stress in treated cells.
Conclusions:
- The combination of NCTD and 2-DG demonstrates potent anti-cancer activity in vitro against liver cancer cells.
- This synergistic effect is mediated through apoptosis induction and oxidative stress.
- Further preclinical investigations are warranted to establish the therapeutic efficacy of NCTD and 2-DG for hepatocellular carcinoma.
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