Investigation and experimental validation of curcumin-related mechanisms against hepatocellular carcinoma based on
Yang Chen1, Qian Li1, Sisi Ren1
1College of Basic Medicine, Shaanxi University of Chinese Medicine, Xianyang 712046, China.
Objectives:
To determine the potential molecular mechanisms underlying the therapeutic effect of curcumin on hepatocellular carcinoma (HCC) by network pharmacology and experimental in vitro validation.
Methods:
The predictive targets of curcumin or HCC were collected from several databases. the identified overlapping targets were crossed with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) platform. Two of the candidate pathways were selected to conduct an experimental verification. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium (MTT) assay was used to determine the effect of curcumin on the viability of HepG2 and LO2 cells. The apoptosis and autophagy of HepG2 cells were respectively detected by flow cytometry and transmission electron microscopy. Besides, western blot and real-time polymerase chain reaction (PCR) were employed to verify the p53 apoptotic pathway and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) autophagy pathway. HepG2 cells were pretreated with pifithrin-α (PFT-α) and GSK690693 for further investigation.
Results:
The 167 pathways analyzed by KEGG included apoptosis, autophagy, p53, and AMPK pathways. The GO enrichment analysis demonstrated that curcumin was involved in cellular response to drug, regulation of apoptotic pathway, and so on. The in vitro experiments also confirmed that curcumin can inhibit the growth of HepG2 cells by promoting the apoptosis of p53 pathway and autophagy through the AMPK pathway. Furthermore, the protein and messenger RNA (mRNA) of the two pathways were downregulated in the inhibitor-pretreated group compared with the experimental group. The damage-regulated autophagy modulator (DRAM) in the PFT-α-pretreated group was downregulated, and p62 in the GSK690693-pretreated group was upregulated.
Conclusions:
Curcumin can treat HCC through the p53 apoptotic pathway and the AMPK/Unc-51-like kinase 1 (ULK1) autophagy pathway, in which the mutual transformation of autophagy and apoptosis may occur through DRAM and p62.
Insights
Curcumin effectively treats hepatocellular carcinoma (HCC) by activating the p53 apoptotic pathway and the adenosine 5'-monophosphate (AMP) activated protein kinase (AMPK) autophagy pathway. These pathways involve the interplay of damage-regulated autophagy modulator (DRAM) and p62, suggesting a mechanism for mutual transformation between apoptosis and autophagy in HCC treatment.
Area of Science:
- Molecular Biology
- Pharmacology
- Oncology
Background:
- Hepatocellular carcinoma (HCC) is a major global health concern.
- Curcumin, a natural compound, exhibits potential anti-cancer properties.
- Understanding the molecular mechanisms of curcumin's therapeutic effects in HCC is crucial.
Purpose of the Study:
- To elucidate the molecular mechanisms of curcumin's therapeutic effects on HCC.
- To validate the role of specific molecular pathways using in vitro experiments.
- To identify key targets and pathways involved in curcumin's action against HCC.
Main Methods:
- Network pharmacology approach to identify predictive targets of curcumin and HCC.
- Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses.
- In vitro validation using MTT assay, flow cytometry, transmission electron microscopy, western blot, and real-time PCR on HepG2 and LO2 cells.
Main Results:
- KEGG analysis identified 167 pathways, including apoptosis, autophagy, p53, and AMPK.
- Curcumin was found to inhibit HepG2 cell growth by promoting p53-mediated apoptosis and AMPK-mediated autophagy.
- Experimental validation confirmed the involvement of p53 and AMPK pathways, with observed changes in DRAM and p62 expression.
Conclusions:
- Curcumin exerts therapeutic effects on HCC via the p53 apoptotic pathway and the AMPK/Unc-51-like kinase 1 (ULK1) autophagy pathway.
- The study suggests a potential mutual transformation between autophagy and apoptosis mediated by DRAM and p62.
- These findings provide a molecular basis for curcumin's application in HCC treatment.
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