Nelfinavir inhibits regulated intramembrane proteolysis of sterol regulatory element binding protein-1 and activating transcription factor 6 in castration-resistant prostate cancer.

Abstract

Nelfinavir induces apoptosis in liposarcoma by inhibiting site-2 protease (S2P) activity, which leads to suppression of regulated intramembrane proteolysis. We postulate similar effects in castration-resistant prostate cancer because it exhibits a lipogenic phenotype. Nelfinavir inhibited androgen receptor activation in androgen-sensitive prostate cancer and the nuclear translocation of the fusion proteins sterol regulatory element binding protein-1 (SREBP-1)-enhanced green fluorescence protein (EGFP) and activating transcription factor 6 (ATF6)-EGFP in castration-resistant prostate cancer cells, viewed under confocal microscopy. Nelfinavir and site-1 protease (S1P) and S2P small interfering RNAs (siRNAs) reduced the proliferation of castration-resistant prostate cancer and induced apoptosis, which was opposed by autophagy. Inhibition of autophagy with hydroxychloroquine was additive to the apoptotic effect of nelfinavir. Western blotting of S1P and S2P siRNA knockdown and/or nelfinavir-treated cells confirmed the accumulation of precursor SREBP-1 and ATF6. 3,4-Dichloroisocoumarin, an S1P inhibitor, did not affect SREBP-1 processing. In contrast, 1,10-phenanthroline, an S2P inhibitor, reproduced the nelfinavir-treated molecular and biological phenotype. Nelfinavir-mediated inhibition of regulated intramembrane proteolysis led to the accumulation of unprocessed SREBP-1 and ATF6. This resulted in sequential endoplasmic reticulum stress, inhibition of the unfolded protein response, reduced fatty acid synthase expression and apoptosis, which was countered by autophagy. Inhibition of autophagy was at least additive to this pro-apoptotic effect. These findings provide new insights into nelfinavir-induced endoplasmic reticulum stress and cancer cell death, and lead us to propose investigating its clinical activity in castration-resistant prostate cancer. This report validates S2P as a therapeutic target in castration-resistant prostate cancer.

• Authors: Min Guan, Kristen Fousek, Warren A Chow
• Journal: The FEBS journal (FEBS J) Vol. 279 Issue 13 Pg. 2399-411 (Jul 2012) ISSN: 1742-4658 [Electronic] England
• PMID: 22540830 (Publication Type: Journal Article)
• Copyright: © 2012 City of Hope and Beckman Research Institute. Journal compilation © 2012 FEBS.

Chemical References

• AR protein, human
• ATF6 protein, human
• Activating Transcription Factor 6
• HIV Protease Inhibitors
• RNA, Messenger
• RNA, Small Interfering
• Receptors, Androgen
• Recombinant Fusion Proteins
• SREBF1 protein, human
• Sterol Regulatory Element Binding Protein 1
• enhanced green fluorescent protein
• Green Fluorescent Proteins
• FASN protein, human
• Fatty Acid Synthase, Type I
• Proprotein Convertases
• Serine Endopeptidases
• membrane-bound transcription factor peptidase, site 1
• Caspases
• Metalloendopeptidases
• MBTPS2 protein, human
• Nelfinavir

Topics

• Activating Transcription Factor 6 (antagonists & inhibitors, genetics, metabolism)
• Apoptosis (drug effects)
• Autophagy
• Blotting, Western
• Caspases (metabolism)
• Castration
• Cell Nucleus (drug effects, metabolism)
• Cell Proliferation (drug effects)
• Endoplasmic Reticulum Stress (drug effects)
• Fatty Acid Synthase, Type I (genetics, metabolism)
• Green Fluorescent Proteins (genetics, metabolism)
• HIV Protease Inhibitors (pharmacology)
• Humans
• Immunoenzyme Techniques
• Male
• Metalloendopeptidases (antagonists & inhibitors, genetics, metabolism)
• Nelfinavir (pharmacology)
• Proprotein Convertases (antagonists & inhibitors, genetics, metabolism)
• Prostatic Neoplasms (genetics, metabolism, pathology)
• Protein Transport
• Proteolysis
• RNA, Messenger (genetics)
• RNA, Small Interfering (genetics)
• Real-Time Polymerase Chain Reaction
• Receptors, Androgen (genetics, metabolism)
• Recombinant Fusion Proteins (genetics, metabolism)
• Reverse Transcriptase Polymerase Chain Reaction
• Serine Endopeptidases (genetics, metabolism)
• Sterol Regulatory Element Binding Protein 1 (antagonists & inhibitors, genetics, metabolism)
• Transcription, Genetic
• Tumor Cells, Cultured