Maria Saveria Gilardini Montani1, Marisa Granato1, Claudio Santoni2, Paola Del Porto3, Nicolò Merendino2, Gabriella D'Orazi4,5, Alberto Faggioni6, Mara Cirone7. 1. Department of Experimental Medicine, La Sapienza University of Rome, V.le Regina Elena 324, 00161, Rome, Italy. 2. Department of Ecological and Biological Sciences (DEB), Tuscia University, Viterbo, Italy. 3. Department of Biology and Biotechnology 'Charles Darwin', La Sapienza University of Rome, Rome, Italy. 4. Department of Research, Advanced Diagnostic and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy. 5. Department of Medical Sciences, Tumor Biology Unit, University "G. D'Annunzio", Chieti, Italy. 6. Department of Experimental Medicine, La Sapienza University of Rome, V.le Regina Elena 324, 00161, Rome, Italy. alberto.faggioni@uniroma1.it. 7. Department of Experimental Medicine, La Sapienza University of Rome, V.le Regina Elena 324, 00161, Rome, Italy. mara.cirone@uniroma1.it.
Abstract
PURPOSE: Histone deacetylase inhibitors (HDACi) are anti-neoplastic agents that are known to affect the growth of different cancer types, but their underlying mechanisms are still incompletely understood. Here, we compared the effects of two HDACi, i.e., Trichostatin A (TSA) and Valproic Acid (VPA), on the induction of cell death and autophagy in pancreatic cancer-derived cells that exhibit a high metastatic capacity and carry KRAS/p53 double mutations. METHODS: Cell viability and proliferation tests were carried out using Trypan blue dye exclusion, MTT and BrdU assays. FACS analyses were carried out to assess cell cycle progression, apoptosis, reactive oxygen species (ROS) production and mitochondrial depolarization, while Western blot and immunoprecipitation analyses were employed to detect proteins involved in apoptosis and autophagy. RESULTS: We found that both VPA and TSA can induce apoptosis in Panc1 and PaCa44 pancreatic cancer-derived cells by triggering mitochondrial membrane depolarization, Cytochrome c release and Caspase 3 activation, although VPA was more effective than TSA, especially in Panc1 cells. As underlying molecular events, we found that ERK1/2 was de-phosphorylated and that the c-Myc and mutant p53 protein levels were reduced after VPA and, to a lesser extent, after TSA treatment. Up-regulation of p21 and Puma was also observed, concomitantly with mutant p53 degradation. In addition, we found that in both cell lines VPA increased the pro-apoptotic Bim level, reduced the anti-apoptotic Mcl-1 level and increased ROS production and autophagy, while TSA was able to induce these effects only in PaCA44 cells. CONCLUSIONS: From our results we conclude that both VPA and TSA can induce pancreatic cancer cell apoptosis and autophagy. VPA appears have a stronger and broader cytotoxic effect than TSA and, thus, may represent a better choice for anti-pancreatic cancer therapy.
PURPOSE: Histone deacetylase inhibitors (HDACi) are anti-neoplastic agents that are known to affect the growth of different cancer types, but their underlying mechanisms are still incompletely understood. Here, we compared the effects of two HDACi, i.e., Trichostatin A (TSA) and Valproic Acid (VPA), on the induction of cell death and autophagy in pancreatic cancer-derived cells that exhibit a high metastatic capacity and carry KRAS/p53 double mutations. METHODS: Cell viability and proliferation tests were carried out using Trypan blue dye exclusion, MTT and BrdU assays. FACS analyses were carried out to assess cell cycle progression, apoptosis, reactive oxygen species (ROS) production and mitochondrial depolarization, while Western blot and immunoprecipitation analyses were employed to detect proteins involved in apoptosis and autophagy. RESULTS: We found that both VPA and TSA can induce apoptosis in Panc1 and PaCa44 pancreatic cancer-derived cells by triggering mitochondrial membrane depolarization, Cytochrome c release and Caspase 3 activation, although VPA was more effective than TSA, especially in Panc1 cells. As underlying molecular events, we found that ERK1/2 was de-phosphorylated and that the c-Myc and mutant p53 protein levels were reduced after VPA and, to a lesser extent, after TSA treatment. Up-regulation of p21 and Puma was also observed, concomitantly with mutant p53 degradation. In addition, we found that in both cell lines VPA increased the pro-apoptotic Bim level, reduced the anti-apoptotic Mcl-1 level and increased ROS production and autophagy, while TSA was able to induce these effects only in PaCA44 cells. CONCLUSIONS: From our results we conclude that both VPA and TSA can induce pancreatic cancer cell apoptosis and autophagy. VPA appears have a stronger and broader cytotoxic effect than TSA and, thus, may represent a better choice for anti-pancreatic cancer therapy.
Authors: M Göttlicher; S Minucci; P Zhu; O H Krämer; A Schimpf; S Giavara; J P Sleeman; F Lo Coco; C Nervi; P G Pelicci; T Heinzel Journal: EMBO J Date: 2001-12-17 Impact factor: 11.598
Authors: Mara Cirone; Livia Di Renzo; Lavinia Vittoria Lotti; Valeria Conte; Pankaj Trivedi; Roberta Santarelli; Roberta Gonnella; Luigi Frati; Alberto Faggioni Journal: PLoS One Date: 2012-03-07 Impact factor: 3.240
Authors: M Granato; V Lacconi; M Peddis; L V Lotti; L Di Renzo; L D Renzo; R Gonnella; R Santarelli; P Trivedi; L Frati; G D'Orazi; A Faggioni; M Cirone Journal: Cell Death Dis Date: 2013-07-18 Impact factor: 8.469
Authors: Renata L Markman; Liana P Webber; Carlos H V Nascimento Filho; Leonardo A Reis; Pablo A Vargas; Marcio A Lopes; Virgilio Zanella; Manoela D Martins; Cristiane H Squarize; Rogerio M Castilho Journal: Cell Oncol (Dordr) Date: 2018-12-11 Impact factor: 6.730
Authors: Marisa Granato; Maria Saveria Gilardini Montani; Roberta Santarelli; Gabriella D'Orazi; Alberto Faggioni; Mara Cirone Journal: J Exp Clin Cancer Res Date: 2017-11-28