SILAC-Based Mass Spectrometry Analysis Reveals That Epibrassinolide Induces Apoptosis via Activating Endoplasmic Reticulum Stress in Prostate Cancer Cells.
Pinar Obakan,Carlos A. Barrero,Ajda Coker-Gurkan,Elif Damla Arisan,Salim Merali,Narcin Palavan-Unsal +5 more
TLDR
It is suggested that EBR promotes ER stress and induces apoptosis and downregulated calnexin and upregulated BiP and IRE1α expression levels and induced CHOP translocation from the cytoplasm to nucleus, which confirmed the alteration of the ER pathway due to drug treatment.Abstract:
Epibrassinolide (EBR) is a polyhydroxylated sterol derivative and biologically active compound of the brassinosteroids. In addition to well-described roles in plant growth, EBR induces apoptosis in the LNCaP prostate cancer cells expressing functional androgen receptor (AR). Therefore, it is suggested that EBR might have an inhibitory potential on androgen receptor signaling pathway. However, the mechanism by which EBR exerts its effects on LNCaP is poorly understood. To address this gap in knowledge, we used an unbiased global proteomics approach, i.e., stable-isotope labeling by amino acids in cell culture (SILAC). In total, 964 unique proteins were identified, 160 of which were differentially expressed after 12 h of EBR treatment. The quantification of the differentially expressed proteins revealed that the expression of the unfolded protein response (UPR) chaperone protein, calreticulin (CALR), was dramatically downregulated. The decrease in CALR expression was also validated by immunoblotting. Because our data revealed the involvement of the UPR in response to EBR exposure, we evaluated the expression of the other UPR proteins. We demonstrated that EBR treatment downregulated calnexin and upregulated BiP and IRE1α expression levels and induced CHOP translocation from the cytoplasm to nucleus. The translocation of CHOP was associated with caspase-9 and caspase-3 activation after a 12 h EBR treatment. Co-treatment of EBR with rapamycin, an upstream mTOR pathway inhibitor, prevented EBR-induced cell viability loss and PARP cleavage in LNCaP prostate cancer cells, suggesting that EBR could induce ER stress in these cells. In addition, we observed similar results in DU145 cells with nonfunctional androgen receptor. When proteasomal degradation of proteins was blocked by MG132 co-treatment, EBR treatment further induced PARP cleavage relative to drug treatment alone. EBR also induced Ca2+ sequestration, which confirmed the alteration of the ER pathway due to drug treatment. Therefore, we suggest that EBR promotes ER stress and induces apoptosis.read more
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Anti-Cancer Agents in Proliferation and Cell Death: The Calcium Connection
Elizabeth Varghese,Samson Mathews Samuel,Zuhair Sadiq,Peter Kubatka,Alena Liskova,Jozef Benacka,Peter Pazinka,Peter Kruzliak,Dietrich Büsselberg +8 more
TL;DR: How calcium signaling is targeted by anti-cancer drugs is discussed and the role of calcium signaling in epigenetic modification and the Warburg effect in tumorigenesis is highlighted.
Journal ArticleDOI
Therapeutic Potential of Brassinosteroids in Biomedical and Clinical Research
Sukhmeen Kaur Kohli,Abhay Bhardwaj,Vinay Bhardwaj,Anket Sharma,Namarta Kalia,Marco Landi,Renu Bhardwaj +6 more
TL;DR: This review is an attempt to update the information about prospects of Brassinosteroids in biomedical and clinical application.
Journal ArticleDOI
Calreticulin is a fine tuning molecule in epibrassinolide-induced apoptosis through activating endoplasmic reticulum stress in colon cancer cells.
Pınar Obakan-Yerlikaya,Elif Damla Arisan,Ajda Coker-Gurkan,Kaan Adacan,Utku Ozbey,Berna Somuncu,Didem Baran,Narcin Palavan-Unsal +7 more
TL;DR: This study revealed that EBR treatment caused ER stress and UPR by altering CALR expression causing caspase‐dependent apoptosis in HCT 116, HT29, DLD‐1, and SW480 colon cancer cells, while 48 h EBRtreatment did not caused UPR in Fetal Human Colon cells (FHC) and Mouse Embryonic Fibroblast cells (MEF).
Journal ArticleDOI
Flow-cytometric analysis of reactive oxygen species in cancer cells under treatment with brassinosteroids.
Pyotr Kisselev,Olesya V. Panibrat,Aliaksei R. Sysa,Marina V. Anisovich,Vladimir N. Zhabinskii,Vladimir A. Khripach +5 more
TL;DR: Brassinosteroids induced generation of reactive oxygen species (ROS) in A549 cells and their growth in a time and dose‐dependent manner and the maximal effect was observed for (22S,23S)‐28‐homocastasterone which at 30 &mgr;M concentration showed a 6‐fold increase of ROS generation in comparison with the control.
Journal ArticleDOI
Stable Isotope Labeling with Amino Acids (SILAC)-Based Proteomics of Primary Human Kidney Cells Reveals a Novel Link between Male Sex Hormones and Impaired Energy Metabolism in Diabetic Kidney Disease.
Sergi Clotet,Sergi Clotet,María José Soler,Marta Riera,Julio Pascual,Fei Fang,Joyce Zhou,Ihor Batruch,Stella K. Vasiliou,Stella K. Vasiliou,Apostolos Dimitromanolakis,Clara Barrios,Eleftherios P. Diamandis,James W. Scholey,James W. Scholey,Ana Konvalinka,Ana Konvalinka +16 more
TL;DR: This in depth quantitative proteomics study of human primary PTEC response to sex hormone administration suggests that male sex hormone stimulation results in perturbed energy metabolism in kidney cells, and that this perturbation results in increased oxidative stress in the renal cortex.
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