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Nimbolide Exhibits Potent Anticancer Activity Through ROS-Mediated ER Stress and DNA Damage in Human Non-small Cell Lung Cancer Cells

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Abstract

The non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancers. It is usually diagnosed at an advanced stage with poor prognosis. Nimbolide (NB), a terpenoid limonoid isolated from the flowers and leaves of neem tree, possesses anticancer properties in various cancer cell lines. However, the underlying mechanism of its anticancer effect on human NSCLC cells remains unclear. In the present study, we investigated the effect of NB on A549 human NSCLC cells. We found that NB treatment inhibits A549 cells colony formation in a dose-dependent manner. Mechanistically, NB treatment increases cellular reactive oxygen species (ROS) level, leading to endoplasmic reticulum (ER) stress, DNA damage, and eventually induction of apoptosis in NSCLC cells. Furthermore, all these effects of NB were blocked by pretreatment with antioxidant glutathione (GSH), the specific ROS inhibitor. We further knockdown CHOP protein by siRNA markedly reduced NB-induced apoptosis in A549 cells. Taken together, our findings reveal that NB is an inducer of ER stress and ROS; these findings may contribute to increasing the therapeutic efficiency of NSCLC.

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Abbreviations

53BP1:

Tumor protein p53 binding protein 1

ATF4:

Activating transcription factor 4

Bax:

Bcl2-associated protein x

Bcl2:

B-cell lymphoma 2

DCFH-DA:

2′,7′-Dichlorodihydrofluorescein diacetate

DSB:

Double-strand break

EGFR:

Epidermal growth factor receptor

ER:

Endoplasmic reticulum

FITC:

Fluorescein isothiocyanate

GSH:

Glutathione

HRP:

Horseradish peroxidase

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NB:

Nimbolide

NSCLC:

Non-small cell lung cancer

PI:

Propidium iodide

ROS:

Reactive oxygen species

STAT3:

Signal transducer and activator of transcription 3

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Funding

This study was supported by the Medical Health Science and Technology Project of Zhejiang Provincial Health Commission (No. 2021KY399) and the Science and Technology Plan Project of Taizhou (No. 22ywb83).

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Author notes

  1. Xi Chen, Hangshuo Zhang, and Yuzhu Pan contribute equally to this work.

Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, Taizhou University, Jiaojiang, Taizhou, 318000, Zhejiang, China

    Xi Chen & Guang Chen

  2. Department of Clinical Medicine, School of Medicine, Taizhou University, Jiaojiang, Taizhou, 318000, Zhejiang, China

    Hangshuo Zhang & Yuzhu Pan

  3. Municipal Hospital Affiliated to Taizhou University, Jiaojiang, Taizhou, 318000, Zhejiang, China

    Ning Zhu & Jiabing Wang

  4. Department of Biochemistry, School of Medicine, Taizhou University, Jiaojiang, Taizhou, 318000, Zhejiang, China

    Lisha Zhou

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Contributions

This work was carried out in collaboration among all authors. X C, JB W, and N Z contributed to the literature search and study design. X C, JB W, and G C participated in the drafting of the article. X C, N Z, HS Z, and YZ P carried out the experiments. JB W and LS Z revised the manuscript. IB W, HS Z, and YZ P contributed to data collection and analysis. All authors have read and approved the final manuscript.

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Correspondence to Xi Chen or Jiabing Wang.

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Chen, X., Zhang, H., Pan, Y. et al. Nimbolide Exhibits Potent Anticancer Activity Through ROS-Mediated ER Stress and DNA Damage in Human Non-small Cell Lung Cancer Cells. Appl Biochem Biotechnol 196, 182–202 (2024). https://doi.org/10.1007/s12010-023-04507-9

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