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LFHP-1c Attenuates Hepatocellular Carcinoma Viability In Vitro Independent of PGAM5
Ganesan Muthusamy1, Chin-Chi Liu2, Andrea N Johnston1,3
1School of Veterinary Medicine, Veterinary Clinical Sciences, Louisiana State University, Baton Rouge, LA 70802, USA.
Background/Objectives:
Upregulation of phosphoglycerate mutase 5 (PGAM5) is correlated with reduced survival outcomes in hepatocellular carcinoma (HCC). PGAM5 knockdown or knockout attenuates HCC growth in in vitro and in vivo models. A novel small molecule inhibitor of PGAM5, LFHP-1c, has recently been characterized. The objective of this study was to determine if LFHP-1c effectively reduces HCC viability in cell models.
Methods:
The hepatoma and HCC cell lines, HepG2 and HuH7, respectively, were treated with LFHP-1c. Label-free imaging was used to quantify growth. Cellular viability and reactive oxygen species (ROS) production were measured using luminescent or fluorescent assays. Expression of antioxidant and metabolic proteins was measured by immunoblot. HepG2 and HuH7 PGAM5 knockout cell lines were used as negative controls.
Results:
Treatment with LFHP-1c reduced cell growth and viability in HepG2 and HuH7 cell lines. Reactive oxygen species production was upregulated in both wild-type and PGAM5 knockout cell lines following LFHP-1c exposure. Cell viability was reduced following LFHP-1c treatment in PGAM5 knockout cell lines.
Conclusions:
LFHP-1c reduces hepatoma and HCC viability and enhances ROS production, but these effects are independent of PGAM5.
Insights
The small molecule LFHP-1c reduces hepatocellular carcinoma (HCC) cell viability and increases reactive oxygen species (ROS). These effects occur independently of phosphoglycerate mutase 5 (PGAM5) inhibition.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Phosphoglycerate mutase 5 (PGAM5) is upregulated in hepatocellular carcinoma (HCC), correlating with poor survival.
- Inhibiting PGAM5 has shown potential in reducing HCC growth in preclinical models.
- A novel small molecule inhibitor, LFHP-1c, targeting PGAM5 has been developed.
Purpose of the Study:
- To evaluate the efficacy of LFHP-1c in reducing HCC cell viability.
- To investigate the mechanism of action of LFHP-1c in hepatoma and HCC cell lines.
Main Methods:
- HepG2 and HuH7 cell lines were treated with LFHP-1c.
- Cell growth was quantified using label-free imaging.
- Cell viability and reactive oxygen species (ROS) production were measured via luminescence and fluorescence assays.
- Protein expression of antioxidant and metabolic factors was assessed using immunoblotting.
- PGAM5 knockout cell lines served as controls.
Main Results:
- LFHP-1c treatment decreased cell growth and viability in both HepG2 and HuH7 cell lines.
- LFHP-1c exposure led to increased ROS production in both wild-type and PGAM5 knockout cells.
- Cell viability was reduced by LFHP-1c even in PGAM5 knockout cells.
Conclusions:
- LFHP-1c effectively reduces hepatoma and HCC cell viability.
- LFHP-1c enhances ROS production in HCC cells.
- The observed effects of LFHP-1c are independent of PGAM5.
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