Icariin plus curcumol enhances autophagy through the mTOR pathway and promotes cathepsin B-mediated pyroptosis of prostate cancer cells

Abstract

Objective: To examine the effect of icariin plus curcumol on prostate cancer cells PC3 and elucidate the underlying mechanisms. Methods: We employed the Cell Counting Kit 8 assay and colony formation assay to assess cell viability and proliferation. Autophagy expression was analyzed using monodansylcadaverine staining. Immunofluorescence and Western blot analyses were used to evaluate protein expressions related to autophagy, pyroptosis, and the mTOR pathway. Cellular damage was examined using the lactate dehydrogenase assay. Moreover, cathepsin B and NLRP3 were detected by co-immunoprecipitation. Results: Icariin plus curcumol led to a decrease in PC3 cell proliferation and an enhancement of autophagy. The levels of LC3-Ⅱ/LC3-Ⅰ and beclin-1 were increased, while the levels of p62 and mTOR were decreased after treatment with icariin plus curcumol. These changes were reversed upon overexpression of mTOR. Furthermore, 3-methyladenine resulted in a decrease in inflammatory cytokines, pyroptosis-related protein levels, and lactate dehydrogenase concentration, compared to the icariin plus curcumol group. Inhibiting cathepsin B reversed the regulatory effects of icariin plus curcumol. Conclusions: Icariin plus curcumol demonstrates great potential as a therapeutic agent for castration-resistant prostate cancer by enhancing autophagy via the mTOR pathway and promoting pyroptosis mediated by cathepsin B. These findings provide valuable insights into the molecular mechanisms underlying the therapeutic potential of icariin and curcumol for prostate cancer treatment.

Keywords

Icariin / Curcumol / Autophagy / mTOR / Cathepsin B / Pyroptosis / Prostate cancer

Conflict of interest statement

The authors declare that they have no conflict of interest.

Funding

This study was supported by Natural Science Foundation of Hunan Province (No. 2023JJ40511), Excellent Youth Project of Scientific Research Program of Hunan Education Department (No. 22B0370), Project of Traditional Chinese Medicine Administration of Hunan Province (No. B2023034), Science and Technology Development Foundation of Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University (No. LYYB202214), and Hunan Provincial Hygiene and Health Commission Health Research Project (No. W20243165).

Data availability statement

The data supporting the findings of this study are available from the corresponding authors upon request.

Authors’ contributions

WS supervised, conceived and designed the study; XYW and WJX searched the literature. XYW, WJX, BNL, and TSS conducted the experiments and acquired the data. All authors contributed to data and statistical analyses. XYW and WJX prepared, WS edited, and all authors reviewed the manuscript. All authors have accepted responsibility for the entire content of this manuscript and approved its publication.

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