2. Academic Validation
  3. Hypericin photoactivation induces triple-negative breast cancer cells pyroptosis by targeting the ROS/CALR/Caspase-3/GSDME pathway

Hypericin photoactivation induces triple-negative breast cancer cells pyroptosis by targeting the ROS/CALR/Caspase-3/GSDME pathway

  • J Adv Res. 2025 Jan 25:S2090-1232(25)00059-1. doi: 10.1016/j.jare.2025.01.042.
Chen Liang 1,  Xiao Liu 2,  Jie Yu 2,  Lingyun Shi 2,  Wenchao Wei 2,  Yalu Zhu 2,  Maoping Feng 2,  Tingting Tang 1,  Dameng Li 1,  Tao Yang 1,  Junnian Zheng 3,  Bo Ma 4,  Liang Wei 5
Affiliations

Affiliations

  • 1 Cancer Institute, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China; Center of Clinical Oncology, The Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou 221002 Jiangsu, China; Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China.
  • 2 Cancer Institute, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China.
  • 3 Center of Clinical Oncology, The Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou 221002 Jiangsu, China; Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China. Electronic address: jnzheng@xzhmu.edu.cn.
  • 4 Cancer Institute, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China; Center of Clinical Oncology, The Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou 221002 Jiangsu, China; Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China. Electronic address: boma@xzhmu.edu.cn.
  • 5 Cancer Institute, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China; Center of Clinical Oncology, The Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou 221002 Jiangsu, China; Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004 Jiangsu, China. Electronic address: weiliang@xzhmu.edu.cn.
PMID: 39870303 DOI: 10.1016/j.jare.2025.01.042
Abstract

Introduction: Hypericin (HP), a natural photosensitizer, has demonstrated great efficacy in photodynamic therapy (PDT) for Cancer treatment. In addition to the induction of Apoptosis and necrosis through Reactive Oxygen Species (ROS) generation, the therapeutic mechanisms and targets of PDT-HP remain unknown.

Objectives: To investigate the direct targets and mechanisms of action of photoactivated hypericin in the inhibition of triple-negative breast Cancer (TNBC).

Methods: Cell Pyroptosis was examined via LDH release, SYTOX Green staining, and ELISA. RNA Sequencing, network pharmacology, drug affinity target stability (DARTS)-tandem mass spectrometry (MS/MS), and molecular docking were employed to identify drug targets. Furthermore, immunoblotting and flow cytometry were utilized to elucidate the mechanisms of drug action.

Results: Our research revealed that PDT-HP can induce Pyroptosis in TNBC cells. Further investigation revealed that PDT-HP induces endoplasmic reticulum stress, activating Caspase-3 and gasdermin E (GSDME) to trigger TNBC cell Pyroptosis. RNA-seq, network pharmacology, and DARTS-MS/MS proteomic analyses revealed that the endoplasmic reticulum protein calreticulin (CALR) is a potential HP target and that interfering with CALR inhibited PDT-HP-induced Pyroptosis. During PDT-HP treatment, the interaction between CALR and SERCA2 inactivates SERCA2, increasing the susceptibility of cells to increased intracellular CA2+ levels under oxidative stress. This triggered endoplasmic reticulum stress and activated Caspase3, which further cleaved GSDME, releasing GSDME-N and ultimately leading to Pyroptosis in TNBC cells.

Conclusion: In this study, we provide insight into the antitumor mechanism by examining the pharmacological mechanism by which PDT-HP regulates TNBC cell Pyroptosis via the ROS/CALR/Caspase-3/GSDME signaling axis.

Keywords

Calreticulin; Endoplasmic reticulum stress; Hypericin; Photodynamic therapy; Pyroptosis; TNBC.

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