Planta Med 2022; 88(13): 1233-1244 DOI: 10.1055/a-1676-4307
Natural Product Chemistry and Analytical Studies
Original Papers
Quantitative Proteomics Based on iTRAQ Reveal that Nitidine Chloride
Induces Apoptosis by Activating JNK/c-Jun Signaling in Hepatocellular Carcinoma
Cells
Shipeng Chen
1
School of Pharmaceutical Sciences, Guangxi Medical University, Nanning,
Guangxi, China;
2
Department of Pharmacy, Liuzhou Peopleʼs Hospital, Liuzhou, Guangxi,
China
,
Yinan Liao
1
School of Pharmaceutical Sciences, Guangxi Medical University, Nanning,
Guangxi, China;
,
Jinyan Lv
1
School of Pharmaceutical Sciences, Guangxi Medical University, Nanning,
Guangxi, China;
,
Huaxin Hou
1
School of Pharmaceutical Sciences, Guangxi Medical University, Nanning,
Guangxi, China;
1
School of Pharmaceutical Sciences, Guangxi Medical University, Nanning,
Guangxi, China;
› Author AffiliationsSupported by:
National
Natural Science Foundation of China
81960706
Supported by:
Guangxi First-class Discipline Project for Pharmaceutical
Sciences
GXFCDP-PS-2018
Supported by:
Natural Science Foundation of Guangxi Province of
China
2017GXNSFAA198304
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.
1H and 13C NMR data and spectra of NC (Fig. 1S
and Fig. 2S), HR-ESI-MS spectrum of NC (Fig. 3S), distribution of
NC in Bel-7402 cells (Fig. 4S), protein distributions detected by
proteomics (Fig. 5S), repeated experiments performed for obtaining the CV
distribution (Fig. 6S) and cytotoxic effects of NC on three HCC cell
lines (Fig. 7S) are provided as Supporting Information.
Georg Thieme Verlag KG Rüdigerstraße 14, 70469 Stuttgart, Germany
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