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<p>The endoplasmic reticulum (ER) chaperone heat shock protein family A member 5 (HSPA5/GRP78/BiP) regulates the unfolded protein response (UPR). It is aberrantly expressed in breast cancer (BC) and contributes to tumor growth, survival, drug resistance, and metastasis. Therefore, it could be a promising target for BC treatment. This study comprehensively analyzed HSPA5 expression across BC subtypes and investigated the effects of the combination of sanguinarine (SAN) and doxorubicin (DOX) on its activity and expression in breast cancer cells. We extensively analyzed HSPA5 expression in BC subtypes via multiple publicly available databases and examined the impact of HSPA5 expression and genomic alterations on patient survival patterns. Analysis of data from GEO datasets revealed that SAN and DOX treatments downregulated key pathways and genes associated with metastasis including HSPA5 and modulated ICD-related genes in MCF-7 cells. Furthermore, the interaction of SAN with DOX, individually and in combination with the ATP-binding domain of HSPA5, was evaluated via in silico docking and molecular dynamics (MD) simulations. The effects of SAN and DOX and their combination on HSPA5 activity and expression were subsequently assessed via biochemical and ELISA-based assays in MCF-7 cells. Our findings indicate that compared with its normal counterpart, HSPA5 is aberrantly overexpressed in primary and metastatic BC tissues. Elevated HSPA5 expression and its genetic and epigenetic alterations are correlated with reduced overall survival (OS) in BC patients. Docking and MD simulation studies revealed strong and stable interactions among SAN, DOX, and their combination with HSPA5. Biochemical and ELISA-based experimental results demonstrated that SAN and DOX significantly inhibited HSPA5-ATPase activity, similar to HSPA5 inhibitor (HA15) in MCF-7 cells, with IC₅₀ values of 8.6 µM, 6.7 µM, and 3.02 µM, respectively. At their respective IC₅₀ and half-IC₅₀ concentrations, SAN, DOX, and their combination reduced total and cell surface HSPA5 expression by more than 50%, concurrently decreasing MCF-7 cell viability and damaging the integrity of the cell membrane. This combination caused late apoptosis by inducing severe ER stress and excessive ROS. This study offers new insights into the combined effects of SAN and DOX on HSPA5 in BC.</p>

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Sanguinarine and Doxorubicin Inhibit Heat Shock Protein HSPA5 and Trigger ER Stress in Breast Cancer

  • 期刊论文

  • 发表日期：

    2025年10月14日

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  • Priyamvada Bhamidipati,
  • Rama Rao Malla

摘要

The endoplasmic reticulum (ER) chaperone heat shock protein family A member 5 (HSPA5/GRP78/BiP) regulates the unfolded protein response (UPR). It is aberrantly expressed in breast cancer (BC) and contributes to tumor growth, survival, drug resistance, and metastasis. Therefore, it could be a promising target for BC treatment. This study comprehensively analyzed HSPA5 expression across BC subtypes and investigated the effects of the combination of sanguinarine (SAN) and doxorubicin (DOX) on its activity and expression in breast cancer cells. We extensively analyzed HSPA5 expression in BC subtypes via multiple publicly available databases and examined the impact of HSPA5 expression and genomic alterations on patient survival patterns. Analysis of data from GEO datasets revealed that SAN and DOX treatments downregulated key pathways and genes associated with metastasis including HSPA5 and modulated ICD-related genes in MCF-7 cells. Furthermore, the interaction of SAN with DOX, individually and in combination with the ATP-binding domain of HSPA5, was evaluated via in silico docking and molecular dynamics (MD) simulations. The effects of SAN and DOX and their combination on HSPA5 activity and expression were subsequently assessed via biochemical and ELISA-based assays in MCF-7 cells. Our findings indicate that compared with its normal counterpart, HSPA5 is aberrantly overexpressed in primary and metastatic BC tissues. Elevated HSPA5 expression and its genetic and epigenetic alterations are correlated with reduced overall survival (OS) in BC patients. Docking and MD simulation studies revealed strong and stable interactions among SAN, DOX, and their combination with HSPA5. Biochemical and ELISA-based experimental results demonstrated that SAN and DOX significantly inhibited HSPA5-ATPase activity, similar to HSPA5 inhibitor (HA15) in MCF-7 cells, with IC₅₀ values of 8.6 µM, 6.7 µM, and 3.02 µM, respectively. At their respective IC₅₀ and half-IC₅₀ concentrations, SAN, DOX, and their combination reduced total and cell surface HSPA5 expression by more than 50%, concurrently decreasing MCF-7 cell viability and damaging the integrity of the cell membrane. This combination caused late apoptosis by inducing severe ER stress and excessive ROS. This study offers new insights into the combined effects of SAN and DOX on HSPA5 in BC.