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Induction of apoptosis via proteasome inhibition in leukemia/lymphoma cells by two potent piperidones

Cellular Oncology volume 41, pages 623–636 (2018)Cite this article

Abstract

Purpose

Previously, compounds containing a piperidone structure have been shown to be highly cytotoxic to cancer cells. Recently, we found that the piperidone compound P2 exhibits a potent anti-neoplastic activity against human breast cancer-derived cells. Here, we aimed to evaluate two piperidone compounds, P1 and P2, for their potential anti-neoplastic activity against human leukemia/lymphoma-derived cells.

Methods

Cytotoxicity and apoptosis induction were evaluated using MTS, annexin V-FITC/PI and mitochondrial membrane potential polychromatic assays to confirm the mode of action of the piperidone compounds. The effects of compound P1 and P2 treatment on gene expression were assessed using AmpliSeq analysis and, subsequently, confirmed by RT-qPCR and Western blotting.

Results

We found that the two related piperidone compounds P1 and P2 selectively killed the leukemia/lymphoma cells tested at nanomolar concentrations through induction of the intrinsic apoptotic pathway, as demonstrated by mitochondrial depolarization and caspase-3 activation. AmpliSeq-based transcriptome analyses of the effects of compounds P1 and P2 on HL-60 acute leukemia cells revealed a differential expression of hundreds of genes, 358 of which were found to be affected by both. Additional pathway analyses revealed that a significant number of the common genes were related to the unfolded protein response, implying a possible role of the two compounds in the induction of proteotoxic stress. Subsequent analyses of the transcriptome data revealed that P1 and P2 induced similar gene expression alterations as other well-known proteasome inhibitors. Finally, we found that Noxa, an important mediator of the activity of proteasome inhibitors, was significantly upregulated at both the mRNA and protein levels, indicating a possible role in the cytotoxic mechanism induced by P1 and P2.

Conclusions

Our data indicate that the cytotoxic activity of P1 and P2 on leukemia/lymphoma cells is mediated by proteasome inhibition, leading to activation of pro-apoptotic pathways.

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Acknowledgments

Funding for this work was provided by the National Institute of General Medical Sciences-Support of Competitive Research (SCORE) grant 1SC3GM103713 to RJA, as well as a Canadian Institutes of Health Research-Regional Partnerships Program Saskatchewan grant to JRD. We thank Madan Balal for the compound docking experiments presented in the supplementary material. The synthesis and docking work performed by the Dr. Rachid Skouta research laboratory was partially supported by Lung Cancer Research Foundation and Green Fund grants. The authors also thank the Genomic Analysis and Cytometry, Screening and Imaging Core Facilities at the University of Texas at El Paso (UTEP), which were supported by a Research Centers in Minority Institutions (RCMI) program grant 5G12MD007592 to the Border Biomedical Research Center (BBRC) in UTEP from the National Institute on Minority Health and Health Disparities, a component of the National Institutes of Health. The authors thank Gladys Almodovar (with UTEP) for cell culture expertise. LC was supported by NIGMS RISE training grant R25 GM069621-15.

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Author notes

  1. Lisett Contreras and Ruben I. Calderon contributed equally as first authors

Affiliations

  1. Department of Biological Sciences and Border Biomedical Research Center, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX, 79968-0519, USA

    Lisett Contreras, Ruben I. Calderon, Armando Varela-Ramirez & Renato J. Aguilera

  2. Hubei Key Laboratory of Agricultural Bioinformatics, College of Informatics, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Hong-Yu Zhang & Yuan Quan

  3. Drug Discovery and Development Research Group, College of Pharmacy and Nutrition, University of Saskatchewan, Saskatoon, S7N 5E5, Canada

    Umashankar Das & Jonathan R. Dimmock

  4. Department of Chemistry, Border Biomedical Research Center, The University of Texas at El Paso, 500 West University Avenue, El Paso, TX, 79968-0519, USA

    Rachid Skouta

  5. Department of Biology, University of Massachusetts, Amherst, MA, 01003-9297, USA

    Rachid Skouta

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  1. Lisett Contreras
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  2. Ruben I. Calderon
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Correspondence to Renato J. Aguilera.

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Contreras, L., Calderon, R.I., Varela-Ramirez, A. et al. Induction of apoptosis via proteasome inhibition in leukemia/lymphoma cells by two potent piperidones. Cell Oncol. 41, 623–636 (2018). https://doi.org/10.1007/s13402-018-0397-1

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Keywords

  • Anti-cancer drug discovery. Apoptosis. Caspase-3. Mitochondrial depolarization. Proteotoxic stress. Proteasome inhibition. Noxa. Piperidone