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Quantitative Proteomics Based on iTRAQ Reveal that Nitidine Chloride Induces Apoptosis by Activating JNK/c-Jun
Shipeng Chen1,2, Yinan Liao1, Jinyan Lv1
1School of Pharmaceutical Sciences, Guangxi Medical University, Nanning, Guangxi, China.
Abstract:
The aim of the present study was to investigate the cytotoxic effects and underlying molecular mechanisms of nitidine chloride (NC) in hepatocellular carcinoma cells via quantitative proteomics. MTT assays were used to detect the inhibitory effects of NC in Bel-7402 liver cancer cells, and the number of apoptotic cells was measured by flow cytometry. Quantitative proteomics technology based on iTRAQ was used to discover differential expressed proteins after NC treatment, and bioinformatic techniques were further used to screen potential targets of NC. Molecular docking was applied to evaluate the docking activity of NC with possible upstream proteins, and their expression was detected at the mRNA and protein levels by quantitative reverse transcription PCR and western blotting. NC inhibited the proliferation of Bel-7402 cells after 24 h of treatment and stimulated apoptosis in vitro. The proteomics experiment showed that NC triggers mitochondrial damage in HCC cells and transcription factor AP-1 (c-Jun) may be a potential target of NC (fold change = 4.36 ± 0.23). Molecular docking results revealed the highest docking score of NC with c-Jun N-terminal kinase (JNK), one of the upstream proteins of c-Jun. Moreover, the mRNA and protein expression of c-Jun and JNK were significantly increased after NC treatment (p < 0.05). These findings indicate that NC significantly induced mitochondrial damage in HCC cells, and induced apoptosis by activating JNK/c-Jun signaling.
Insights
Nitidine chloride (NC) inhibits liver cancer cell proliferation and induces apoptosis by damaging mitochondria. This occurs through the activation of the JNK/c-Jun signaling pathway, revealing a novel therapeutic mechanism for hepatocellular carcinoma.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Hepatocellular carcinoma (HCC) remains a significant global health challenge.
- Identifying novel therapeutic agents and understanding their mechanisms of action are crucial for improving HCC treatment outcomes.
Purpose of the Study:
- To investigate the cytotoxic effects of nitidine chloride (NC) on hepatocellular carcinoma cells.
- To elucidate the molecular mechanisms underlying NC's action using quantitative proteomics.
Main Methods:
- MTT assays and flow cytometry for proliferation and apoptosis analysis.
- iTRAQ-based quantitative proteomics to identify differentially expressed proteins.
- Bioinformatic analysis, molecular docking, qRT-PCR, and Western blotting to validate targets.
Main Results:
- NC inhibited Bel-7402 liver cancer cell proliferation and induced apoptosis.
- Proteomics identified mitochondrial damage and implicated AP-1 (c-Jun) as a potential target.
- Molecular docking and expression analysis confirmed JNK/c-Jun pathway activation.
Conclusions:
- Nitidine chloride induces hepatocellular carcinoma cell apoptosis via mitochondrial damage.
- The JNK/c-Jun signaling pathway is activated by NC, presenting a potential therapeutic strategy for HCC.
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