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Repurposing fluphenazine to suppress melanoma brain, lung and bone metastasis by inducing G0/G1 cell cycle arrest and apoptosis and disrupting autophagic flux

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Abstract

Brain metastasis is the main cause of treatment failure and melanoma-related death. Inadequate concentrations of therapeutic drugs in the brain due to the blood-brain barrier (BBB) pose a major challenge in the treatment of brain metastasis. Antipsychotics can cross the BBB to reach the brain. Fluphenazine (FPZ) inhibits the survival of melanoma cells in vitro. However, its efficacy in suppressing the metastasis of melanoma, especially brain metastasis, remains unknown. Therefore, we explored whether fluphenazine (FPZ) can be repurposed for treating melanoma metastasis. A subcutaneous tumor model, and experimental metastasis models that simulate the outgrowth of melanoma cells in the brain, lung, and bone were established to verify the inhibitory effect of FPZ on melanoma cells. FPZ showed potential inhibitory effects against melanoma both in vivo and in vitro. It induced G0/G1 phase arrest and-mitochondrion-mediated intrinsic apoptosis, and inhibited autophagic flux in melanoma cells in vitro. In vivo, subcutaneous tumor, brain, lung, and bone models of metastatic melanoma were established. Intraperitoneal injection of FPZ (8 mg/kg) significantly inhibited melanoma growth in the subcutaneous and experimental metastasis models. In a lung metastasis model, FPZ reduced the proportion of M2 macrophages and increased the proportion of CD8+ T cells and NK cells in vivo, thereby promoting an anticancer immune response. The findings of this study indicate that FPZ is a potential drug candidate for treating metastatic melanoma.

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Data availability

All the original data used in our study can available upon request for noncommercial purposes.

Abbreviations

BMs:

brain metastases

MAP:

mitogen-activated protein

BBB:

blood-brain barrier

IC50:

half maximal inhibitory concentration

FCM:

flow cytometry;

ΔΨm:

mitochondrial membrane potential

ROS:

reactive oxygen species;

H&E:

hematoxylin and eosin

CDK:

cyclin-dependent kinase;

p21:

cyclin-dependent kinase inhibitor 1 A

p27:

cyclin-dependent kinase inhibitor 1B

PRAP:

poly ADP-ribose polymerase

Bcl2:

B-cell lymphoma 2;p62/SQSTM1,sequestosome 1

CQ:

chloroquine diphosphate

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Acknowledgements

Thanks to Liyuan Fang (Core Facilities of West China Hospital) for her contribution in the identification of some human melanoma cells lines.

Funding

This work was funded by the National Natural Science Foundation of China (grant No. 82173280), the Department of Science and Technology of Sichuan Province (grant number: 2021YJ0450), the Chengdu Science and Technology Bureau International Cooperation Project (grant number: 2019-GH02-00036-HZ) and Central Government Funds of Guiding Local Scientific and Technological Development for Sichuan Province (grant number: 2021ZYD0070).

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Authors and Affiliations

  1. Department of Rehabilitation Medicine, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, China

    Huizhi Xi, Mengling Wu, Qianrui Huang, Yiwen Zhang & Yong Xia

  2. West China School of Pharmacy, Sichuan University, Chengdu, 60041, China

    Hongbo Ma & Shanrui Li

  3. Key Laboratory of Rehabilitation Medicine in Sichuan Province/Rehabilitation Medicine Research Institute, Chengdu, 610041, China

    Yong Xia

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  1. Huizhi Xi
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Contributions

YX, HZX: Research ideas and experimental design. All authors: Acquisition and interpretation of the data. WML, SRL: Statistical analysis. HZX, QRH: Drafting of the manuscript, HBM: Manuscript revision. YX: Obtained funding. YX, YWZ: Study supervision.

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Correspondence to Yiwen Zhang or Yong Xia.

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The research protocol was approved by the Animal Care and Use Committee of Sichuan University, the written informed consents of all participants was obtained.

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Xi, H., Wu, M., Ma, H. et al. Repurposing fluphenazine to suppress melanoma brain, lung and bone metastasis by inducing G0/G1 cell cycle arrest and apoptosis and disrupting autophagic flux. Clin Exp Metastasis 40, 161–175 (2023). https://doi.org/10.1007/s10585-023-10202-0

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