Lisett Contreras1, Ruben I Calderon1, Armando Varela-Ramirez1, Hong-Yu Zhang2, Yuan Quan2, Umashankar Das3, Jonathan R Dimmock3, Rachid Skouta4,5, Renato J Aguilera6. 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. 2. Hubei Key Laboratory of Agricultural Bioinformatics, College of Informatics, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China. 3. Drug Discovery and Development Research Group, College of Pharmacy and Nutrition, University of Saskatchewan, Saskatoon, S7N 5E5, Canada. 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. 5. Department of Biology, University of Massachusetts, Amherst, MA, 01003-9297, USA. 6. 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. raguilera@utep.edu.
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.
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 humanbreast cancer-derived cells. Here, we aimed to evaluate two piperidone compounds, P1 and P2, for their potential anti-neoplastic activity against humanleukemia/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.
Authors: Paulina J Villanueva; Alberto Martinez; Sarah T Baca; Rebecca E DeJesus; Manuel Larragoity; Lisett Contreras; Denisse A Gutierrez; Armando Varela-Ramirez; Renato J Aguilera Journal: PLoS One Date: 2018-11-05 Impact factor: 3.240
Authors: Lisett Contreras; Stephanie Medina; Austre Y Schiaffino Bustamante; Edgar A Borrego; Carlos A Valenzuela; Umashankar Das; Subhas S Karki; Jonathan R Dimmock; Renato J Aguilera Journal: Pharmacol Rep Date: 2021-08-26 Impact factor: 3.024
Authors: Kinjal Lakhani; Edgar A Borrego; Karla G Cano; Jonathan R Dimmock; Renato J Aguilera; Swagatika Das; Praveen K Roayapalley; Rajendra K Sharma; Umashankar Das Journal: Medicines (Basel) Date: 2021-06-03
Authors: Praveen K Roayapalley; Hiroshi Sakagami; Keitaro Satoh; Shigeru Amano; Kenjiro Bandow; Renato J Aguilera; Karla G Cano Hernandez; Austre Y Schiaffino Bustamante; Stephen G Dimmock; Rajendra K Sharma; Umashankar Das; Jonathan R Dimmock Journal: Medicines (Basel) Date: 2021-12-16