Histone deacetylase inhibitors upregulate Rap1GAP and inhibit Rap activity in thyroid tumor cells

Increases in Rap activity have been associated with tumor progression. Although activating mutations in Rap have not been described, downregulation of Rap1GAP is frequent in human tumors including thyroid carcinomas. In this study, we explored whether endogenous Rap1GAP expression could be restored to thyroid tumor cells. The effects of deacetylase inhibitors and a demethylating agent, individually and in combination, were examined in four differentiated and six anaplastic thyroid carcinoma (ATC) cell lines. Treatment with the structurally distinct histone deacetylase (HDAC) inhibitors, sodium butyrate and trichostatin A, increased Rap1GAP expression in all the differentiated thyroid carcinoma cell lines and in four of the six ATC cell lines. The demethylating agent, 5-aza-deoxycytidine, restored Rap1GAP expression in one anaplastic cell line and enhanced the effects of HDAC inhibitors in a second anaplastic cell line. Western blotting indicated that Rap2 was highly expressed in human thyroid cancer cells. Importantly, treatment with HDAC inhibitors impaired Rap2 activity in both differentiated and anaplastic tumor cell lines. The mechanism through which Rap activity is repressed appears to entail effects on the expression of multiple Rap regulators, including RapGEFs and RapGAPs. These results suggest that HDAC inhibitors may provide a tractable approach to impair Rap activity in human tumor cells.

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Histone deacetylase inhibitors valproate and trichostatin A are toxic to neuroblastoma cells and modulate cytochrome P450 1A1, 1B1 and 3A4 expression in these cells

Interdisciplinary Toxicology ◽

10.2478/v10102-009-0019-x ◽

2009 ◽

Vol 2 (3) ◽

pp. 205-210 ◽

Cited By ~ 15

Author(s):

Jana Hřebačková ◽

Jitka Poljaková ◽

Tomáš Eckschlager ◽

Jan Hraběta ◽

Pavel Procházka ◽

...

Keyword(s):

Cytochrome P450 ◽

Cell Line ◽

Histone Deacetylase ◽

Cell Lines ◽

Histone Deacetylase Inhibitors ◽

Trichostatin A ◽

Neuroblastoma Cell ◽

Neuroblastoma Cells ◽

Dependent Manner ◽

Cytochrome P450 1A1

Histone deacetylase inhibitors valproate and trichostatin A are toxic to neuroblastoma cells and modulate cytochrome P450 1A1, 1B1 and 3A4 expression in these cellsHistone deacetylase inhibitors such as valproic acid (VPA) and trichostatin A (TSA) were shown to exert antitumor activity. Here, the toxicity of both drugs to human neuroblastoma cell lines was investigated using MTT test, and IC50 values for both compounds were determined. Another target of this work was to evaluate the effects of both drugs on expression of cytochrome P450 (CYP) 1A1, 1B1 and 3A4 enzymes, which are known to be expressed in neuroblastoma cells. A malignant subset of neuroblastoma cells, so-called N-type cells (UKF-NB-3 cells) and the more benign S-type neuroblastoma cells (UKF-NB-4 and SK-N-AS cell lines) were studied from both two points of view. VPA and TSA inhibited the growth of neuroblastoma cells in a dose-dependent manner. The IC50values ranging from 1.0 to 2.8 mM and from 69.8 to 129.4 nM were found for VPA and TSA, respectively. Of the neuroblastoma tested here, the N-type UKF-NB-3 cell line was the most sensitive to both drugs. The different effects of VPA and TSA were found on expression of CYP1A1, 1B1 and 3A4 enzymes in individual neuroblastoma cells tested in the study. Protein expression of all these CYP enzymes in the S-type SK-N-AS cell line was not influenced by either of studied drugs. On the contrary, in another S-type cell line, UKF-NB-4, VPA and TSA induced expression of CYP1A1, depressed levels of CYP1B1 and had no effect on expression levels of CYP3A4 enzyme. In the N-type UKF-NB-3 cell line, the expression of CYP1A1 was strongly induced, while that of CYP1B1 depressed by VPA and TSA. VPA also induced the expression of CYP3A4 in this neuroblastoma cell line.

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ABCG2/BCRP gene expression is related to epithelial–mesenchymal transition inducer genes in a papillary thyroid carcinoma cell line (TPC-1)

Journal of Molecular Endocrinology ◽

10.1530/jme-14-0051 ◽

2014 ◽

Vol 52 (3) ◽

pp. 289-300 ◽

Cited By ~ 9

Author(s):

E Mato ◽

C González ◽

A Moral ◽

J I Pérez ◽

O Bell ◽

...

Keyword(s):

Thyroid Carcinoma ◽

Cell Line ◽

Epithelial Mesenchymal Transition ◽

Carcinoma Cell ◽

Thyroid Tumor ◽

Carcinoma Cell Line ◽

Human Thyroid ◽

Mesenchymal Transition ◽

Starting Point ◽

E Cadherin

Tumor malignancy is associated with the epithelial–mesenchymal transition (EMT) process and resistance to chemotherapy. However, little is known about the relationship between the EMT and the multidrug-resistance gene in thyroid tumor progression. We investigated whether the expression of theABCG2/BCRPgene is associated withZEB1and other EMT inducer genes involved in tumor dedifferentiation. We established a subpopulation of cells that express theABCG2/BCRPgene derived from the thyroid papillary carcinoma cell line (TPC-1), the so-called TPC-1 MITO-resistant subline. The most relevant findings in these TPC-1 selected cells were a statistically significant upregulation ofZEB1andTWIST1(35- and 15-fold change respectively), no changes in the relative expression of vimentin andSNAIL1, and no expression of E-cadherin. The TPC-1 MITO-resistant subline displayed a faster migration and greater invasive ability than parental cells in correlation with a significant upregulation of the survivin (BIRC5) gene (twofold change,P<0.05). The knockdown ofZEB1promoted nuclear re-expression of E-cadherin, reduced expression of vimentin, N-cadherin, andBIRC5genes, and reduced cell migration (P<0.05). Analysis of human thyroid carcinoma showed a slight overexpression of theABCG2/BCRPat stages I and II (P<0.01), and a higher overexpression at stages III and IV (P<0.01).SNAIL1,TWIST1, andZEB1genes showed higher expression at stages III and IV than at stages I and II. E- and N-cadherin genes were upregulated at stages I and II of the disease (ninefold and tenfold change, respectively,P<0.01) but downregulated at stages III and IV (fourfold lower,P<0.01). These results could be a promising starting point for further study of the role of theABCG2/BCRPgene in the progression of thyroid tumor.

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Histone Deacetylase Inhibitors and Microtubule Inhibitors Induce Apoptosis in Feline Luminal Mammary Carcinoma Cells

Animals ◽

10.3390/ani11020502 ◽

2021 ◽

Vol 11 (2) ◽

pp. 502

Author(s):

Filipe Almeida ◽

Andreia Gameiro ◽

Jorge Correia ◽

Fernando Ferreira

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Cell Line ◽

Cell Lines ◽

Mammary Carcinoma ◽

Human Breast Cancer ◽

Histone Deacetylase Inhibitors ◽

Trichostatin A ◽

Human Breast ◽

Antitumor Effects

Feline mammary carcinoma (FMC) is the third most common type of neoplasia in cats, sharing similar epidemiological features with human breast cancer. In humans, histone deacetylases (HDACs) play an important role in the regulation of gene expression, with HDAC inhibitors (HDACis) disrupting gene expression and leading to cell death. In parallel, microtubules inhibitors (MTIs) interfere with the polymerization of microtubules, leading to cell cycle arrest and apoptosis. Although HDACis and MTIs are used in human cancer patients, in cats, data is scarce. In this study, we evaluated the antitumor properties of six HDACis (CI-994, panobinostat, SAHA, SBHA, scriptaid, and trichostatin A) and four MTIs (colchicine, nocodazole, paclitaxel, and vinblastine) using three FMC cell lines (CAT-MT, FMCp, and FMCm), and compared with the human breast cancer cell line (SK-BR-3). HDACis and MTIs exhibited dose-dependent antitumor effects in FMC cell lines, and for all inhibitors, the IC50 values were determined, with one feline cell line showing reduced susceptibility (FMCm). Immunoblot analysis confirmed an increase in the acetylation status of core histone protein HDAC3 and flow cytometry showed that HDACis and MTIs lead to cellular apoptosis. Overall, our study uncovers HDACis and MTIs as promising anti-cancer agents to treat FMCs.

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Histone deacetylase inhibitor panobinostat induces antitumor activity in epithelioid sarcoma and rhabdoid tumor by growth factor receptor modulation

BMC Cancer ◽

10.1186/s12885-021-08579-w ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Anne Catherine Harttrampf ◽

Maria Eugenia Marques da Costa ◽

Aline Renoult ◽

Estelle Daudigeos-Dubus ◽

Birgit Geoerger

Keyword(s):

Growth Factor ◽

Histone Deacetylase ◽

Cell Lines ◽

Epithelioid Sarcoma ◽

Hdac Inhibitors ◽

Growth Factor Receptor ◽

Rhabdoid Tumor ◽

Tumor Growth Inhibition ◽

Mesenchymal Transition ◽

Receptor Modulation

Abstract Background Epithelioid sarcomas and rhabdoid tumors are rare, aggressive malignancies with poor prognosis. Both are characterized by INI1 alterations and deregulation of growth factor receptors albeit their interaction has not been elucidated. Methods In this study, we investigated the activity of a panel of epigenetic modulators and receptor tyrosine kinase inhibitors in vitro on respective cell lines as well as on primary patient-derived epithelioid sarcoma cells, and in vivo on xenografted mice. Focusing on histone deacetylase (HDAC) inhibitors, we studied the mechanism of action of this class of agents, its effect on growth factor receptor regulation, and changes in epithelial-to-mesenchymal transition by using cell- and RT-qPCR-based assays. Results Pan-HDAC inhibitor panobinostat exhibited potent anti-proliferative activity at low nanomolar concentrations in A204 rhabdoid tumor, and VAESBJ/GRU1 epithelioid sarcoma cell lines, strongly induced apoptosis, and resulted in significant tumor growth inhibition in VAESBJ xenografts. It differentially regulated EGFR, FGFR1 and FGFR2, leading to downregulation of EGFR in epithelioid sarcoma and to mesenchymal-to-epithelial transition whereas in rhabdoid tumor cells, EGFR was strongly upregulated and reinforced the mesenchymal phenotype. All three cell lines were rendered more susceptible towards combination with EGFF inhibitor erlotinib, further enhancing apoptosis. Conclusions HDAC inhibitors exhibit significant anticancer activity due to their multifaceted actions on cytotoxicity, differentiation and drug sensitization. Our data suggest that the tailored, tissue-specific combination of HDAC inhibitors with therapeutics which target cellular salvage mechanisms might increase their therapeutic relevance.

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Status and expression of thep16INK4 gene in human thyroid tumors and thyroid-tumor cell lines

International Journal of Cancer ◽

10.1002/(sici)1097-0215(19960703)67:1<29::aid-ijc7>3.0.co;2-1 ◽

1996 ◽

Vol 67 (1) ◽

pp. 29-34 ◽

Cited By ~ 18

Author(s):

Viola Calabr� ◽

Maria Strazzullo ◽

Girolama La Mantia ◽

Monica Fedele ◽

Christian Paulin ◽

...

Keyword(s):

Tumor Cell ◽

Cell Lines ◽

Thyroid Tumor ◽

Human Thyroid ◽

Thyroid Tumors ◽

Tumor Cell Lines

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Establishment of Tumor Cell Lines: From Primary Tumor Cells to a Tumor Cell Line

Cell Culture Technology - Learning Materials in Biosciences ◽

10.1007/978-3-319-74854-2_4 ◽

2018 ◽

pp. 61-73

Author(s):

Katharina Meditz ◽

Beate Rinner

Keyword(s):

Tumor Cell ◽

Cell Line ◽

Tumor Cells ◽

Cell Lines ◽

Primary Tumor ◽

Tumor Cell Line ◽

Tumor Cell Lines

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Coinduction of Embryonic and Adult-Type Globin mRNAs by Sodium Butyrate and Trichostatin A in Two Murine Interleukin-3–Dependent Bone Marrow–Derived Cell Lines

Blood ◽

10.1182/blood.v92.11.4383 ◽

1998 ◽

Vol 92 (11) ◽

pp. 4383-4393 ◽

Cited By ~ 4

Author(s):

Kimiko Ishiguro ◽

Alan C. Sartorelli

Keyword(s):

Bone Marrow ◽

Histone Deacetylase ◽

Cell Lines ◽

Sodium Butyrate ◽

Trichostatin A ◽

Globin Genes ◽

Globin Mrna ◽

Interleukin 3 ◽

Mrna Species ◽

Major Species

Abstract Using an RNase protection assay, globin mRNA species expressed in clones derived from Ba/F3 and B6SUtA cells transfected with the erythropoietin receptor (EpoR) and selected with erythropoietin (Epo) were compared with globin mRNA species induced in corresponding parental cells by sodium butyrate (SB) and trichostatin A (TSA). βMajor/βminor- and -1/-2–globin mRNAs were the major species, with trace amounts of ɛ-globin mRNA, formed in Epo-stimulated EpoR+ Ba/F3 clones, whereas SB and TSA allowed expression of all species of globin mRNAs, ie, ɛ, βh1, βmajor/βminor, ζ, and -1/-2, in parental Ba/F3 cells. In contrast, ɛ- and -1/-2–globin mRNAs were the major species present in Epo-stimulated EpoR+ B6SUtA clones, whereas SB and TSA activated ɛ-, βh1-, βS/βT-, and -1/-2–globin genes in parental B6SUtA cells; ζ-globin mRNA was not detected in SB- and TSA-treated B6SUtA cells. Because TSA is a specific inhibitor of histone deacetylase, the mimicry of action exhibited by SB and TSA suggests that the effects of SB are mediated through its ability to inhibit histone deacetylase and that histone deacetylase is an integral part of the repression of globin genes in these interleukin-3–dependent cells. Efficient coinduction of embryonic and adult types of globin mRNA in bone marrow cell lines derived from adult mice indicates that adult hematopoietic precursors possess an embryonic nature. These cell lines are useful models to study the mechanism(s) of developmental globin gene switching.

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Dye-mediated photolysis is capable of eliminating drug-resistant (MDR+) tumor cells

Blood ◽

10.1182/blood.v81.3.793.793 ◽

1993 ◽

Vol 81 (3) ◽

pp. 793-800 ◽

Cited By ~ 21

Author(s):

RM Lemoli ◽

T Igarashi ◽

M Knizewski ◽

L Acaba ◽

A Richter ◽

...

Keyword(s):

Cell Line ◽

Tumor Cells ◽

Cell Lines ◽

Light Exposure ◽

Lymphoblastic Leukemia ◽

Leukemia Cell ◽

Myelogenous Leukemia ◽

Leukemia Cell Line ◽

Drug Resistant ◽

Colony Forming Unit

Abstract We evaluated the potential role of photoradiation therapy with a benzoporphyrin derivative, monoacid ring A (BPD-MA), and dihematoporphyrin ether (DHE), for the ex vivo purging of residual tumor cells from autologous bone marrow (BM) grafts. BPD-MA and DHE photosensitizing activity was tested against two human large-cell lymphoma cell lines and colony-forming unit-leukemia (CFU-L) derived from patients with acute myelogenous leukemia (AML). In mixing experiments, 4-log elimination of tumor cell lines was observed after 1 hour of incubation with 75 ng/mL of BPD-MA or 30 minutes of treatment with 12.5 micrograms/mL of DHE followed by white light exposure. By comparison, using the same concentration of BPD-MA, the mean recovery of normal BM progenitors was 4% +/- 0.8% (mean +/- SD) for granulocyte- macrophage colony-forming unit (CFU-GM) and 5% +/- 0.8% for burst- forming unit-erythroid (BFU-E). Similarly, DHE treatment resulted in the recovery of 5.2% +/- 2% and 9.8% +/- 3% of CFU-GM and BFU-E, respectively. Furthermore, equivalently cytotoxic concentrations of both DHE and BPD-MA and light were found not to kill normal pluripotent stem cells in BM, as demonstrated by their survival in two-step long- term marrow culture at levels equal to untreated controls. The T- lymphoblastic leukemia cell line CEM and its vinblastine (VBL)- resistant subline CEM/VBL, along with the acute promyelocyte leukemia cell line HL-60 and its vincristine (VCR)-resistant subline HL-60/VCR, were also tested. BPD-MA at 75 ng/mL was able to provide a greater than 4-log elimination of the drug-sensitive cell lines, but only a 34% and 55% decrease of the drug-resistant HL-60/VCR and CEM/VBL cell lines, respectively. On the contrary, 12.5 micrograms/mL of DHE reduced the clonogenic growth of all the cell lines by more than 4 logs. Further experiments demonstrated decreased uptake of both BPD-MA and DHE by the resistant cell lines. However, all the cell lines took up more DHE than BPD-MA under similar experimental conditions. Our results demonstrate the preferential cytotoxicity of BPD-MA and DHE toward neoplastic cell lines and CFU-L from AML patients. In addition, DHE was slightly more effective in purging tumor cells expressing the p-170 glycoprotein. These results suggest that photoradiation with DHE would be useful for in vitro purging of residual drug-resistant leukemia and lymphoma cells.

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Histone Deacetylase Inhibitors Down-Regulate bcl-2 Expression and Induce Apoptosis in t(14;18) Lymphomas

Molecular and Cellular Biology ◽

10.1128/mcb.25.5.1608-1619.2005 ◽

2005 ◽

Vol 25 (5) ◽

pp. 1608-1619 ◽

Cited By ~ 184

Author(s):

Hong Duan ◽

Caroline A. Heckman ◽

Linda M. Boxer

Keyword(s):

Cell Cycle ◽

Histone Deacetylase ◽

Chromatin Immunoprecipitation ◽

Histone Deacetylase Inhibitors ◽

Antitumor Agents ◽

Trichostatin A ◽

Hdac Inhibitors ◽

Transcriptional Level ◽

Cycle Arrest ◽

Induced Apoptosis

ABSTRACT Histone deacetylase (HDAC) inhibitors are promising antitumor agents, but they have not been extensively explored in B-cell lymphomas. Many of these lymphomas have the t(14;18) translocation, which results in increased bcl-2 expression and resistance to apoptosis. In this study, we examined the effects of two structurally different HDAC inhibitors, trichostatin A (TSA) and sodium butyrate (NaB), on the cell cycle, apoptosis, and bcl-2 expression in t(14;18) lymphoma cells. We found that in addition to potent cell cycle arrest, TSA and NaB also dramatically induced apoptosis and down-regulated bcl-2 expression, and overexpression of bcl-2 inhibited TSA-induced apoptosis. The repression of bcl-2 by TSA occurred at the transcriptional level. Western blot analysis and quantitative chromatin immunoprecipitation (ChIP) assay showed that even though HDAC inhibitors increased overall acetylation of histones, localized histone H3 deacetylation occurred at both bcl-2 promoters. TSA treatment increased the acetylation of the transcription factors Sp1 and C/EBPα and decreased their binding as well as the binding of CBP and HDAC2 to the bcl-2 promoters. Mutation of Sp1 and C/EBPα binding sites reduced the TSA-induced repression of bcl-2 promoter activity. This study provides a mechanistic rationale for the use of HDAC inhibitors in the treatment of human t(14;18) lymphomas.

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