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1.
BMC Cancer ; 21(1): 493, 2021 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-33941107

RESUMO

BACKGROUND: Glioblastoma (GBM) is the deadliest and the most common primary brain tumor in adults. The invasiveness and proliferation of GBM cells can be decreased through the inhibition of Wnt/ß-catenin pathway. In this regard, celecoxib is a promising agent, but other COXIBs and 2,5-dimethylcelecoxib (2,5-DMC) await elucidation. Thus, the aim of this study was to analyze the impact of celecoxib, 2,5-DMC, etori-, rofe-, and valdecoxib on GBM cell viability and the activity of Wnt/ß-catenin pathway. In addition, the combination of the compounds with temozolomide (TMZ) was also evaluated. Cell cycle distribution and apoptosis, MGMT methylation level, COX-2 and PGE2 EP4 protein levels were also determined in order to better understand the molecular mechanisms exerted by these compounds and to find out which of them can serve best in GBM therapy. METHODS: Celecoxib, 2,5-DMC, etori-, rofe- and valdecoxib were evaluated using three commercially available and two patient-derived GBM cell lines. Cell viability was analyzed using MTT assay, whereas alterations in MGMT methylation level were determined using MS-HRM method. The impact of COXIBs, in the presence and absence of TMZ, on Wnt pathway was measured on the basis of the expression of ß-catenin target genes. Cell cycle distribution and apoptosis analysis were performed using flow cytometry. COX-2 and PGE2 EP4 receptor expression were evaluated using Western blot analysis. RESULTS: Wnt/ß-catenin pathway was attenuated by COXIBs and 2,5-DMC irrespective of the COX-2 expression profile of the treated cells, their MGMT methylation status, or radio/chemoresistance. Celecoxib and 2,5-DMC were the most cytotoxic. Cell cycle distribution was altered, and apoptosis was induced after the treatment with celecoxib, 2,5-DMC, etori- and valdecoxib in T98G cell line. COXIBs and 2,5-DMC did not influence MGMT methylation status, but inhibited COX-2/PGE2/EP4 pathway. CONCLUSIONS: Not only celecoxib, but also 2,5-DMC, etori-, rofe- and valdecoxib should be further investigated as potential good anti-GBM therapeutics.


Assuntos
Neoplasias Encefálicas/metabolismo , Inibidores de Ciclo-Oxigenase 2/farmacologia , Glioblastoma/metabolismo , Proteínas de Neoplasias/efeitos dos fármacos , Pirazóis/farmacologia , Sulfonamidas/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , Idoso , Antineoplásicos Alquilantes/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Encefálicas/tratamento farmacológico , Celecoxib/farmacologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ciclo-Oxigenase 2/metabolismo , Metilases de Modificação do DNA/efeitos dos fármacos , Metilases de Modificação do DNA/metabolismo , Enzimas Reparadoras do DNA/efeitos dos fármacos , Enzimas Reparadoras do DNA/metabolismo , Dinoprostona/metabolismo , Relação Dose-Resposta a Droga , Etoricoxib/farmacologia , Feminino , Glioblastoma/tratamento farmacológico , Humanos , Isoxazóis/farmacologia , Lactonas/farmacologia , Masculino , Metilação , Pessoa de Meia-Idade , Proteínas de Neoplasias/metabolismo , Receptores de Prostaglandina E Subtipo EP4/efeitos dos fármacos , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Sulfonas/farmacologia , Temozolomida/farmacologia , Proteínas Supressoras de Tumor/efeitos dos fármacos , Proteínas Supressoras de Tumor/metabolismo , beta Catenina/efeitos dos fármacos , beta Catenina/metabolismo
2.
Clin Epigenetics ; 13(1): 35, 2021 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-33588926

RESUMO

BACKGROUND: Epigenetic inactivation of O6-methylguanine DNA-methyltransferase (MGMT) is associated with increased sensitivity to alkylating chemotherapeutic agents in glioblastoma patients. The genetic background underlying MGMT gene methylation may explain individual differences in treatment response and provide a clue to a personalized treatment strategy. Making use of the longitudinal twin design, we aimed, for the first time, to estimate the genetic contributions to MGMT methylation in a Danish twin cohort. METHODS: DNA-methylation from whole blood (18 monozygotic (MZ) and 25 dizygotic (DZ) twin pairs) repeated 10 years apart from the Longitudinal Study of Aging Danish Twins (LSADT) were used to search for genetic and environmental contributions to DNA-methylation at 170 CpG sites of across the MGMT gene. Both univariate and bivariate twin models were applied. The intraclass correlations, performed on cross-sectional data (246 MZ twin pairs) from an independent study population, the Middle-Aged Danish Twins (MADT), were used to assess the genetic influence at each CpG site of MGMT for replication. RESULTS: Univariate twin model revealed twelve CpG sites showing significantly high heritability at intake (wave 1, h2 > 0.43), and seven CpG sites with significant heritability estimates at end of follow-up (wave 2, h2 > 0.5). There were six significant CpG sites, located at the gene body region, that overlapped among the two waves (h2 > 0.5), of which five remained significant in the bivariate twin model, which was applied to both waves. Within MZ pair correlation in these six CpGs from MADT demarks top level of genetic influence. There were 11 CpGs constantly have substantial common environmental component over the 10 years. CONCLUSIONS: We have identified 6 CpG sites linked to the MGMT gene with strong and persistent genetic control based on their DNA methylation levels. The genetic basis of MGMT gene methylation could help to explain individual differences in glioblastoma treatment response and most importantly, provide references for mapping the methylation Quantitative Trait Loci (meQTL) underlying the genetic regulation.


Assuntos
Metilases de Modificação do DNA/genética , Enzimas Reparadoras do DNA/genética , DNA/metabolismo , Glioblastoma/genética , Guanina/análogos & derivados , Proteínas Supressoras de Tumor/genética , Idoso , Envelhecimento/genética , Antineoplásicos Alquilantes/uso terapêutico , Estudos de Coortes , Ilhas de CpG/genética , Estudos Transversais , Metilação de DNA , Metilases de Modificação do DNA/efeitos dos fármacos , Enzimas Reparadoras do DNA/efeitos dos fármacos , Dinamarca/epidemiologia , Meio Ambiente , Epigenômica , Feminino , Inativação Gênica/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Guanina/metabolismo , Humanos , Estudos Longitudinais , Masculino , Metiltransferases , Pessoa de Meia-Idade , Proteínas Supressoras de Tumor/efeitos dos fármacos , Gêmeos Dizigóticos/genética , Gêmeos Monozigóticos/genética
3.
CNS Neurosci Ther ; 27(5): 552-563, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33460245

RESUMO

BACKGROUND: Temozolomide (TMZ) is a first-line chemotherapy drug for the treatment of malignant glioma and resistance to it poses a major challenge. Receptor-interacting protein 2 (RIP2) is associated with the malignant character of cancer cells. However, it remains unclear whether RIP2 is involved in TMZ resistance in glioma. METHODS: RIP2 expression was inhibited in TMZ-resistant glioma cells and normal glioma cells by using small interfering RNA (siRNA) against RIP2. Plasmid transfection method was used to overexpress RIP2. Cell counting kit-8 assays were performed to evaluate cell viability. Western blotting or immunofluorescence was performed to determine RIP2, NF-κB, and MGMT expression in cells. Flow cytometry was used to investigate cell apoptosis. TMZ-resistant glioma xenograft models were established to evaluate the role of the RIP2/NF-κB/MGMT signaling pathway in drug resistance. RESULTS: We observed that RIP2 expression was upregulated in TMZ-resistant glioma cells, whereas silencing of RIP2 expression enhanced cellular sensitivity to TMZ. Similarly, upon the induction of RIP2 overexpression, glioma cells developed resistance to TMZ. The molecular mechanism underlying the process indicated that RIP2 can activate the NF-κB signaling pathway and upregulate the expression of O6-methylguanine-DNA methyltransferase (MGMT), following which the glioma cells develop drug resistance. In the TMZ-resistant glioma xenograft model, treatment with JSH-23 (an NF-κB inhibitor) and lomeguatrib (an MGMT inhibitor) could enhance the sensitivity of the transplanted tumor to TMZ. CONCLUSION: We report that the RIP2/NF-κB/MGMT signaling pathway is involved in the regulation of TMZ resistance. Interference with NF-κB or MGMT activity could constitute a novel strategy for the treatment of RIP2-positive TMZ-resistant glioma.


Assuntos
Antineoplásicos Alquilantes/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Metilases de Modificação do DNA/efeitos dos fármacos , Metilases de Modificação do DNA/genética , Enzimas Reparadoras do DNA/efeitos dos fármacos , Enzimas Reparadoras do DNA/genética , Resistencia a Medicamentos Antineoplásicos/genética , Glioma/tratamento farmacológico , NF-kappa B/efeitos dos fármacos , NF-kappa B/genética , Proteína Serina-Treonina Quinase 2 de Interação com Receptor/efeitos dos fármacos , Proteína Serina-Treonina Quinase 2 de Interação com Receptor/genética , Transdução de Sinais/efeitos dos fármacos , Temozolomida/farmacologia , Proteínas Supressoras de Tumor/efeitos dos fármacos , Proteínas Supressoras de Tumor/genética , Animais , Apoptose/efeitos dos fármacos , Neoplasias Encefálicas/genética , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioma/genética , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/antagonistas & inibidores , RNA Interferente Pequeno/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
4.
Nutrients ; 12(3)2020 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-32183060

RESUMO

Breast cancer is a common cancer that occurs due to different epigenetic alterations and genetic mutations. Various epidemiological studies have demonstrated an inverse correlation between breast cancer incidence and flavonoid intake. The anti-cancer action of flavonoids, a class of polyphenolic compounds that are present in plants, as secondary metabolites has been a major topic of research for many years. Our review analysis demonstrates that flavonoids exhibit anti-cancer activity against breast cancer occurring in different ethnic populations. Breast cancer subtype and menopausal status are the key factors in inducing the flavonoid's anti-cancer action in breast cancer. The dose is another key factor, with research showing that approximately 10 mg/day of isoflavones is required to inhibit breast cancer occurrence. In addition, flavonoids also influence the epigenetic machinery in breast cancer, with research demonstrating that epigallocatechin, genistein, and resveratrol all inhibited DNA methyltransferase and altered chromatin modification in breast cancer. These flavonoids can induce the expression of different tumor suppressor genes that may contribute to decreasing breast cancer progression and metastasis. Additional studies are required to confirm the contribution of epigenetic modifications by flavonoids to breast cancer prevention.


Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/genética , Epigênese Genética/efeitos dos fármacos , Flavonoides/farmacologia , Polifenóis/farmacologia , Neoplasias da Mama/prevenção & controle , Neoplasias da Mama/terapia , Catequina/análogos & derivados , Catequina/farmacologia , Cromatina/efeitos dos fármacos , Metilases de Modificação do DNA/efeitos dos fármacos , Feminino , Genes Supressores de Tumor/efeitos dos fármacos , Genisteína/farmacologia , Humanos , Resveratrol/farmacologia
5.
Br J Haematol ; 186(1): 91-100, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30891745

RESUMO

Sickle cell disease (SCD) affects over 2 million people worldwide with high morbidity and mortality in underdeveloped countries. Therapeutic interventions aimed at reactivating fetal haemoglobin (HbF) is an effective approach for improving survival and ameliorating the clinical severity of SCD. A class of agents that inhibit DNA methyltransferase (DNMT) activity show promise as HbF inducers because off-target effects are not observed at low concentrations. However, these compounds are rapidly degraded by cytidine deaminase when taken by oral administration, creating a critical barrier to clinical development for SCD. We previously demonstrated that microRNA29B (MIR29B) inhibits de novo DNMT synthesis, therefore, the goal of our study was to determine if MIR29 mediates HbF induction. Overexpression of MIR29B in human KU812 cells and primary erythroid progenitors significantly increased the percentage of HbF positive cells, while decreasing the expression of DNMT3A and the HBG repressor MYB. Furthermore, HBG promoter methylation levels decreased significantly following MIR29B overexpression in human erythroid progenitors. We subsequently, observed higher MIR29B expression in SCD patients with higher HbF levels compared to those with low HbF. Our findings provide evidence for the ability of MIR29B to induce HbF and supports further investigation to expand treatment options for SCD.


Assuntos
Anemia Falciforme/genética , Epigênese Genética/efeitos dos fármacos , Hemoglobina Fetal/genética , MicroRNAs/fisiologia , Ativação Transcricional/efeitos dos fármacos , gama-Globinas/genética , Linhagem Celular , Células Cultivadas , DNA (Citosina-5-)-Metiltransferases/biossíntese , DNA (Citosina-5-)-Metiltransferases/efeitos dos fármacos , DNA Metiltransferase 3A , Metilases de Modificação do DNA/biossíntese , Metilases de Modificação do DNA/efeitos dos fármacos , Células Precursoras Eritroides/metabolismo , Hemoglobina Fetal/metabolismo , Humanos , MicroRNAs/antagonistas & inibidores
6.
Curr Top Med Chem ; 18(28): 2380-2394, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30430946

RESUMO

Epigenetics process is the heritable change in gene function that does not involve changes in the DNA sequence. Until now, several types of epigenetic mechanisms have been characterized, including DNA methylation, histone modification (acetylation, methylation, etc.), nucleosome remodeling, and noncoding RNAs. With the biological investigations of these modifiers, some of them are identified as promoters in the process of various diseases, such as cancer, cardiovascular disease and virus infection. Epigenetic changes may serve as potential "first hits" for tumorigenesis. Hence, targeting epigenetic modifiers is being considered as a promising way for disease treatment. To date, six agents in two epigenetic target classes (DNMT and HDAC) have been approved by the US Food and Drug Administration (FDA). Most of these drugs are applied in leukemia, lymphoma therapy, or are combined with other drugs for the treatment of solid tumor. Due to the rapid development of epigenetics and epigenetics targeted drugs, it is becoming an emerging area in targeted drug design.


Assuntos
Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Desenho de Fármacos , Epigênese Genética/efeitos dos fármacos , Terapia de Alvo Molecular/métodos , Neoplasias/tratamento farmacológico , Carcinogênese , Metilases de Modificação do DNA/efeitos dos fármacos , Histona Desacetilases/efeitos dos fármacos , Humanos , Neoplasias/enzimologia , Neoplasias/genética
7.
World Neurosurg ; 116: e147-e161, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29709748

RESUMO

BACKGROUND: There is limited information on O6-methylguanine DNA methyltransferase (MGMT) status, extent of surgical resection, and its impact on overall outcomes in patients with glioblastoma (GBM). METHODS: After institutional review board approval, 233 newly diagnosed patients with GBM with known MGMT status (2009-2015) were included in our analysis. Clinical, imaging, and follow-up data were collected from the database. Overall survival (OS) and progression-free survival (PFS) were the primary and secondary end points, respectively. RESULTS: Of patients, 51.9% were younger than 65 years and 44.2% were noted to have promoter methylation of MGMT. Median residual tumor volume was 1.1 cm3 and extent of complete resection of enhancing tumor on imaging was 96%. Estimated median OS and PFS were 10.9 months and 5.4 months, respectively. MGMT status was an independent predictor of PFS (hazard ratio [HR], 0.52; P = 0.005) but only marginally associated with OS (P = 0.059). In MGMT methylated patients, extent of resection (≥86%) and good performance status (Karnofsky Performance Status ≥70) were independently associated with PFS and OS, respectively (PFS: HR, 0.21; P = 0.015; OS: HR, 0.05; P = 0.002). In MGMT promoter unmethylated patients, extent of resection (≥86%) was independently associated with OS (P = 0.039). Concurrent chemoradiotherapy was associated with OS/PFS irrespective of age and MGMT status. CONCLUSIONS: Greater extent of resection of enhancing tumor was associated with improved PFS in MGMT promoter methylated patients, OS regardless of MGMT status. Elderly patients with methylated MGMT promoter were found to have improved PFS whereas younger patients had improved OS with MGMT promoter methylated status.


Assuntos
Antineoplásicos Alquilantes/uso terapêutico , Glioblastoma/patologia , Guanina/análogos & derivados , Metiltransferases/metabolismo , Neoplasia Residual/patologia , Adulto , Idoso , Biomarcadores Tumorais/genética , Neoplasias Encefálicas/cirurgia , Metilases de Modificação do DNA/efeitos dos fármacos , Metilases de Modificação do DNA/metabolismo , Enzimas Reparadoras do DNA/efeitos dos fármacos , Enzimas Reparadoras do DNA/metabolismo , Intervalo Livre de Doença , Feminino , Glioblastoma/genética , Glioblastoma/cirurgia , Guanina/farmacologia , Humanos , Masculino , Pessoa de Meia-Idade
8.
Carcinogenesis ; 39(4): 601-613, 2018 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-29452350

RESUMO

Targeting tumor-initiating, drug-resistant populations of cancer stem cells (CSC) with phytochemicals is a novel paradigm for cancer prevention and treatment. We herein employed a phenotypic drug discovery approach coupled to mechanism-of-action profiling and target deconvolution to identify phenolic components of extra virgin olive oil (EVOO) capable of suppressing the functional traits of CSC in breast cancer (BC). In vitro screening revealed that the secoiridoid decarboxymethyl oleuropein aglycone (DOA) could selectively target subpopulations of epithelial-like, aldehyde dehydrogenase (ALDH)-positive and mesenchymal-like, CD44+CD24-/low CSC. DOA could potently block the formation of multicellular tumorspheres generated from single-founder stem-like cells in a panel of genetically diverse BC models. Pretreatment of BC populations with noncytotoxic doses of DOA dramatically reduced subsequent tumor-forming capacity in vivo. Mice orthotopically injected with CSC-enriched BC-cell populations pretreated with DOA remained tumor-free for several months. Phenotype microarray-based screening pointed to a synergistic interaction of DOA with the mTOR inhibitor rapamycin and the DNA methyltransferase (DNMT) inhibitor 5-azacytidine. In silico computational studies indicated that DOA binds and inhibits the ATP-binding kinase domain site of mTOR and the S-adenosyl-l-methionine (SAM) cofactor-binding pocket of DNMTs. FRET-based Z-LYTE™ and AlphaScreen-based in vitro assays confirmed the ability of DOA to function as an ATP-competitive mTOR inhibitor and to block the SAM-dependent methylation activity of DNMTs. Our systematic in vitro, in vivo and in silico approaches establish the phenol-conjugated oleoside DOA as a dual mTOR/DNMT inhibitor naturally occurring in EVOO that functionally suppresses CSC-like states responsible for maintaining tumor-initiating cell properties within BC populations.


Assuntos
Acetatos/farmacologia , Neoplasias da Mama/patologia , Células-Tronco Neoplásicas/efeitos dos fármacos , Azeite de Oliva/química , Extratos Vegetais/farmacologia , Piranos/farmacologia , Animais , Monoterpenos Ciclopentânicos , Metilases de Modificação do DNA/efeitos dos fármacos , Feminino , Humanos , Camundongos , Serina-Treonina Quinases TOR/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
9.
Neuro Oncol ; 20(2): 236-248, 2018 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-29016925

RESUMO

Background: Glioblastoma (GBM) is almost invariably fatal due to failure of standard therapy. The relapse of GBM following surgery, radiation, and systemic temozolomide (TMZ) is attributed to the ability of glioma stem cells (GSCs) to survive, evolve, and repopulate the tumor mass, events on which therapy exerts a poorly understood influence. Methods: Here we explore the molecular and cellular evolution of TMZ resistance as it emerges in vivo (xenograft models) in a series of human GSCs with either proneural (PN) or mesenchymal (MES) molecular characteristics. Results: We observed that the initial response of GSC-initiated intracranial xenografts to TMZ is eventually replaced by refractory growth pattern. Individual tumors derived from the same isogenic GSC line expressed divergent and complex profiles of TMZ resistance markers, with a minor representation of O6-methylguanine DNA methyltransferase (MGMT) upregulation. In several independent TMZ-resistant tumors originating from MES GSCs we observed a consistent diminution of mesenchymal features, which persisted in cell culture and correlated with increased expression of Nestin, decline in transglutaminase 2 and sensitivity to radiation. The corresponding mRNA expression profiles reflective of TMZ resistance and stem cell phenotype were recapitulated in the transcriptome of exosome-like extracellular vesicles (EVs) released by GSCs into the culture medium. Conclusions: Intrinsic changes in the tumor-initiating cell compartment may include loss of subtype characteristics and reciprocal alterations in sensitivity to chemo- and radiation therapy. These observations suggest that exploiting therapy-induced changes in the GSC phenotype and alternating cycles of therapy may be explored to improve GBM outcomes.


Assuntos
Antineoplásicos Alquilantes/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Vesículas Extracelulares/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Células-Tronco Neoplásicas/efeitos dos fármacos , Temozolomida/farmacologia , Animais , Neoplasias Encefálicas/tratamento farmacológico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Metilases de Modificação do DNA/efeitos dos fármacos , Metilases de Modificação do DNA/genética , Vesículas Extracelulares/metabolismo , Humanos , Camundongos , Células-Tronco Neoplásicas/metabolismo
11.
Toxicology ; 361-362: 62-72, 2016 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-27396502

RESUMO

Mammalian spermatogenesis is a progressive process comprising spermatogonial proliferation, spermatocytic meiosis, and later spermiogenesis, which is considered to be under the regulation of epigenetic parameters. To gain insights into the significance of DNA methylation in early spermatogenesis, 5-azadC was used as a molecular biological tool to mimic the level of DNA methylation in vivo. Since the drug is incorporated into DNA during the S-phase, spermatogonia and spermatocytes would be affected primarily in mouse spermatogenesis. Adult male ICR mice were intraperitoneally injected with 5-azadC at a dose of 0.25mg/kg/day for 10 consecutive days, allowing us to examine its maximum effect on the kinetics of spermatogonia and spermatocytes. In this short-term protocol, 5-azadC induced significant histological abnormalities, such as a marked increase in apoptosis of spermatogonia and spermatocytes, followed by severe loss of spermatids, while after termination of 5-azadC treatment, normal histology was restored in the testis within 35days. Quantification of the methylation level of CCGG sites as well as whole DNA showed spermatogonial hypomethylation, which correlated with increased apoptosis of spermatogonia. Interestingly, the hypomethylated cells were simultaneously positive for tri-methylated histone H3 at K4. On the other hand, no changes in methylation level were found in spermatocytes, but PCNA staining clearly showed disordered accumulation of S-phase spermatocytes, which increased their apoptosis in stage XII. In addition, different immunohistochemical staining pattern was found for DNA methyltransferases (DNMTs); DNMT1was expressed in the majority of all germ cells, but DNMT3a and b were only expressed in spermatogonia. Our results indicate that 5-azadC caused DNA hypomethylation in spermatogonia, but induced prolongation of S-phase in spermatocytes, resulting in the induction of apoptosis in both cases. Thus, 5-azadC affects spermatogenesis at more than one differentiation stage with different mechanisms, probably due to the specific usage of DNMTs.


Assuntos
Antimetabólitos Antineoplásicos/toxicidade , Azacitidina/análogos & derivados , Metilases de Modificação do DNA/metabolismo , Espermatogênese/efeitos dos fármacos , Animais , Azacitidina/toxicidade , Diferenciação Celular/efeitos dos fármacos , Metilação de DNA/efeitos dos fármacos , Metilases de Modificação do DNA/efeitos dos fármacos , Decitabina , Células Germinativas/efeitos dos fármacos , Células Germinativas/ultraestrutura , Masculino , Camundongos , Camundongos Endogâmicos ICR , Túbulos Seminíferos/efeitos dos fármacos , Túbulos Seminíferos/ultraestrutura , Espermatócitos/efeitos dos fármacos , Espermatócitos/metabolismo , Espermatogônias/efeitos dos fármacos , Espermatogônias/metabolismo , Testículo/efeitos dos fármacos , Testículo/metabolismo
12.
Zhongguo Zhong Xi Yi Jie He Za Zhi ; 36(10): 1219-1223, 2016 10.
Artigo em Chinês | MEDLINE | ID: mdl-30641010

RESUMO

Objective To observe the effect of Wenhua Juanbi Recipe (WJR) on the expres- sions of DNA methyltransferases (DNMTs) in peripheral blood mononuclear cells (PBMCs) of collagen- inducing arthritis (CIA) , and to study its mechanism for treating CIA. Methods Totally 90 Wistar rats were randomly divided into the model group (n =80) and the normal control group (n = 10). Rats of the model group were injected with type II collagen of bovine (BC II) emulsion from the tail to establish CIA model. Successfully modeled 50 CIA rats were randomly divided into five groups, i.e., the model group, the methotrexate (MTX) group, the low dose WJR group, the middle dose WJR group, the high dose WJR group, 10 in each group. Rats in the model group were administered with normal saline by gastrogavage, once per day. Rats in low, middle, and high dose WJR groups were administered with WJR by gas- trogavage at the daily dose of 22. 9, 45. 8, 68. 7 g/kg, respectively (once per day). Rats in the MTX group were administered with MTX suspension (0.78 mg/kg) by gastrogavage, once per week for 30 successive days. The paw swelling was evaluated using volume method (draining volume). PBMCs were extrac- ted from each group after intervention. mRNA expression levels of DNMTs (DNMT1 , DNMT3a, DNMT3b) were detected by real-time quantitative PCR. Results Compared with the normal group, the paws were obviously swollen in the model group (P <0. 01). Compared with the model group, swollen paws were obviously alleviated in low, middle, and high dose WJR groups, and the MTX group (P <0.01). Compared with before treatment in the same group, swollen paws were obviously alleviated in low, middle, and high dose WJR groups, and the MTX group (P <0. 01 ). Compared with the normal group, expression levels of DNMT1, DNMT3a, and DNMT3b in PBMCs were obviously lowered in the model group (P <0.01). Compared with the model group, expression levels of DNMT1 , DNMT3a, DNMT3b in PBMCs were obviously elevated in low, middle, and high dose WJR groups, and the MTX group (all P <0. 01). There was no sig- nificant difference in expression levels of DNMT1, DNMT3a, or DNMT3b in PBMCs among low, middle, and high dose WJR groups (P>0.05). Conclusions Expression levels of DNMTs in PBMCs of CIA rats decreased. WJR up-regulated the expression level of DNMTs in PBMCs of CIA rats in no obvious dose de- pendent way. One of WJR's mechanisms for treating CIA might be up-regulating expression levels of DN- MTs, and adjusting the state of DNA methylation.


Assuntos
Artrite , Metilação de DNA , Metilases de Modificação do DNA , Leucócitos Mononucleares , Animais , Artrite/tratamento farmacológico , Artrite/metabolismo , Bovinos , Colágeno , DNA , Metilação de DNA/efeitos dos fármacos , Metilases de Modificação do DNA/efeitos dos fármacos , Metilases de Modificação do DNA/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Ratos , Ratos Wistar
13.
Endocrinology ; 156(12): 4672-83, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26372177

RESUMO

The increasing incidence of reproductive anomalies, described as testicular dysgenesis syndrome, is thought to be related to the exposure of the population to chemicals in the environment. Bisphenol A (BPA) and di(2-ethylhexyl)phthalate (DEHP), which have hormonal and antihormonal activity, have attracted public attention due to their presence in consumer products. The present study investigated the effects of BPA and DEHP on reproductive development. Timed-pregnant female rats were exposed to BPA and DEHP by gavage from gestational days 12 to 21. Results showed that prenatal exposures to test chemicals exerted variable effects on steroidogenic factor 1 and GATA binding protein 4 protein expression and increased (P < .05) sex-determining region Y-box 9 and antimüllerian hormone protein in the infantile rat testis compared with levels in the control unexposed animals. Pituitary LHß and FSHß subunit protein expression was increased (P < .05) in BPA- and DEHP-exposed prepubertal male rats but were decreased (P < .05) in adult animals relative to control. Exposure to both BPA and DEHP in utero inhibited (P < .05) global DNA hydroxymethylation in the adult testis in association with altered DNA methyltransferase protein expression. Together the present data suggest that altered developmental programming in the testes associated with chemical exposures are related to the disruption of sexual differentiation events and DNA methylation patterns. The chemical-induced effects impact the development of steroidogenic capacity in the adult testis.


Assuntos
Compostos Benzidrílicos/farmacologia , Dietilexilftalato/farmacologia , Poluentes Ambientais/farmacologia , Estrogênios não Esteroides/farmacologia , Fenóis/farmacologia , Plastificantes/farmacologia , Diferenciação Sexual/efeitos dos fármacos , Testículo/efeitos dos fármacos , Animais , Hormônio Antimülleriano/metabolismo , Metilação de DNA/efeitos dos fármacos , Metilases de Modificação do DNA/efeitos dos fármacos , Metilases de Modificação do DNA/metabolismo , Disruptores Endócrinos/farmacologia , Feminino , Subunidade beta do Hormônio Folículoestimulante/efeitos dos fármacos , Subunidade beta do Hormônio Folículoestimulante/metabolismo , Fator de Transcrição GATA4/efeitos dos fármacos , Fator de Transcrição GATA4/metabolismo , Disgenesia Gonadal , Hormônio Luteinizante Subunidade beta/efeitos dos fármacos , Hormônio Luteinizante Subunidade beta/metabolismo , Masculino , Hipófise/efeitos dos fármacos , Hipófise/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Ratos , Proteína da Região Y Determinante do Sexo/efeitos dos fármacos , Proteína da Região Y Determinante do Sexo/metabolismo , Fator Esteroidogênico 1/efeitos dos fármacos , Fator Esteroidogênico 1/metabolismo , Doenças Testiculares , Testículo/metabolismo
14.
Neurochem Res ; 37(6): 1192-200, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22228201

RESUMO

Glioblastoma multiforme (GBM) is recognized as the most common and lethal form of central nervous system cancer. Currently used surgical techniques, chemotherapeutic agents, and radiotherapy strategies have done very little in extending the life expectancies of patients diagnosed with GBM. The difficulty in treating this malignant disease lies both in its inherent complexity and numerous mechanisms of drug resistance. In this review, we summarize several of the primary mechanisms of drug resistance. We reviewed available published literature in the English language regarding drug resistance in glioblastoma. The reasons for drug resistance in glioblastoma include drug efflux, hypoxic areas of tumor cells, cancer stem cells, DNA damage repair, and miRNAs. Many potential therapies target these mechanisms, including a series of investigated alternative and plant-derived agents. Future research and clinical trials in glioblastoma patients should pursue combination of therapies to help combat drug resistance. The emerging new data on the potential of plant-derived therapeutics should also be closely considered and further investigated.


Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos , Glioblastoma/tratamento farmacológico , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Transportadores de Cassetes de Ligação de ATP/metabolismo , Compostos Alílicos/uso terapêutico , Inibidores da Angiogênese/uso terapêutico , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Terapia Combinada , Metilases de Modificação do DNA/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Enzimas Reparadoras do DNA/efeitos dos fármacos , Flavonoides/uso terapêutico , Alho/química , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , MicroRNAs/uso terapêutico , Células-Tronco Neoplásicas/patologia , Proteínas Proto-Oncogênicas c-bcl-2/efeitos dos fármacos , RNA Interferente Pequeno/uso terapêutico , Retinoides/uso terapêutico , Sulfetos/uso terapêutico , Proteínas Supressoras de Tumor/efeitos dos fármacos
16.
J Neurooncol ; 92(2): 227-32, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19037596

RESUMO

We report a case of a 51-year-old woman with newly diagnosed glioblastoma multiforme (GBM) who was treated with surgery followed by the standard concomitant temozolomide (TMZ) and radiotherapy (RT). Although TMZ is generally safe and well-tolerated, she developed a sudden onset of prolonged and severe thrombocytopenia as the most prominent event of pancytopenia during the combined treatment, leading to discontinuation of the combined therapy. Thrombocytopenia lasted for more than 2 months with intensive, intermittent platelet transfusions. A bone marrow aspiration and biopsy performed after recovery of severe suppression still revealed reduced number of megakaryocytes. O(6)-methylguanine-DNA methyltransferase (MGMT) analyses showed methylated MGMT promoter in GBM, but unmethylated promoters in both peripheral blood leukocytes and bone marrow cells. This is the first report suggesting the irrelevance of MGMT status of normal hematopoietic cells to TMZ-induced severe thrombocytopenia and pancytopenia.


Assuntos
Neoplasias Encefálicas/terapia , Metilases de Modificação do DNA/efeitos dos fármacos , Enzimas Reparadoras do DNA/efeitos dos fármacos , Glioblastoma/terapia , Pancitopenia/etiologia , Trombocitopenia/etiologia , Proteínas Supressoras de Tumor/efeitos dos fármacos , Antineoplásicos Alquilantes/efeitos adversos , Sequência de Bases , Western Blotting , Neoplasias Encefálicas/patologia , Terapia Combinada , Metilação de DNA/efeitos dos fármacos , Metilases de Modificação do DNA/genética , Enzimas Reparadoras do DNA/genética , Dacarbazina/efeitos adversos , Dacarbazina/análogos & derivados , Feminino , Glioblastoma/patologia , Humanos , Imageamento por Ressonância Magnética , Pessoa de Meia-Idade , Procedimentos Neurocirúrgicos , Reação em Cadeia da Polimerase , Radioterapia/efeitos adversos , Temozolomida , Proteínas Supressoras de Tumor/genética
17.
Cleft Palate Craniofac J ; 45(5): 545-51, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18788878

RESUMO

OBJECTIVES: The aim of this study was to analyze epigenetic (specifically, DNA methylation) participation in the mechanisms of cleft palate only induced by maternal exposure to all-trans retinoic acid in mice. DESIGN: Cleft palate only was induced in fetuses by maternal exposure to all-trans retinoic acid. Their secondary palates were excised for analysis. Cytosine extension assay and restriction landmark genomic scanning were performed to analyze DNA methylation status. The expression levels of the DNA methyltransferases were examined by real-time reverse transcriptase-polymerase chain reaction. RESULTS: Using cytosine extension assay, on gestation day 14.5, the status of DNA methylation within CpG islands and in global DNA was decreased significantly in all-trans retinoic acid-treated groups compared with the controls (p < .01 and p < .05). In the controls, the status within CpG islands on gestation day 14.5 was significantly increased compared with gestation days 13.5 and 18.5 (p < .01). Using real-time reverse transcriptase-polymerase chain reaction, there was no significant change in the expression of DNA methyltransferases, except on gestation day 18.5. Using restriction landmark genomic scanning on gestation day 18.5, five spots (0.49%) in the controls and one spot (0.1%) in all-trans retinoic acid-treated groups were specifically detected. CONCLUSIONS: These results indicate that changes in DNA methylation may play an important role in the manifestation of cleft palate only caused by environmental factors such as maternal exposure to all-trans retinoic acid.


Assuntos
Fissura Palatina/induzido quimicamente , Metilação de DNA/efeitos dos fármacos , Tretinoína/efeitos adversos , Animais , Fissura Palatina/embriologia , Ilhas de CpG/efeitos dos fármacos , Citosina/análise , DNA/análise , DNA/efeitos dos fármacos , Metilases de Modificação do DNA/análise , Metilases de Modificação do DNA/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Feminino , Idade Gestacional , Exposição Materna/efeitos adversos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Palato/efeitos dos fármacos , Palato/embriologia , Mapeamento por Restrição , Reação em Cadeia da Polimerase Via Transcriptase Reversa
18.
Carcinogenesis ; 29(6): 1267-75, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18204074

RESUMO

Nickel (Ni) compounds are potent carcinogens and can induce malignant transformation of rodent and human cells. To uncover the molecular mechanisms of nickel sulfide (NiS)-induced cell transformation, we investigated epigenetic alterations in a set of DNA repair genes. The silencing of the O(6)-methylguanine DNA methyltransferase (MGMT) gene locus and upregulation of DNA methyltransferase 1 (DNMT1) expression was specifically detected in NiS-transformed human bronchial epithelial (16HBE) cells. In addition, we noted epigenetic alterations including DNA hypermethylation, reduced histone H4 acetylation and a decrease in the ratio of Lys-9 acetylated/methylated histone H3 at the MGMT CpG island in NiS-transformed 16HBE cells. Meanwhile, we identified concurrent binding of methyl-CpG-binding protein 2, methylated DNA-binding domain protein 2 and DNMT1 to the CpG island of the MGMT promoter, demonstrating that these components collaborate to maintain MGMT methylation in NiS-transformed cells. Moreover, depletion of DNMT1 by introduction of a small hairpin RNA construct into NiS-transformed cells resulted in a 30% inhibition of cell proliferation and led to increased MGMT gene expression by reversion of the epigenetic modifications at the MGMT promoter region. MGMT suppression and hypermethylation at the CpG island of the MGMT promoter occurred 6 days after NiS treatment, indicating that epigenetic modifications of MGMT might be an early event in tumorigenesis. Taken together, these observations demonstrate that epigenetic silencing of MGMT is associated with DNA hypermethylation, histone modifications and DNMT1 upregulation, which contribute to NiS-induced malignant transformation.


Assuntos
Carcinógenos/toxicidade , Transformação Celular Neoplásica/genética , DNA (Citosina-5-)-Metiltransferases/genética , Metilases de Modificação do DNA/genética , Enzimas Reparadoras do DNA/genética , Inativação Gênica , Níquel/toxicidade , Proteínas Supressoras de Tumor/genética , Western Blotting , Linhagem Celular , DNA (Citosina-5-)-Metiltransferase 1 , DNA (Citosina-5-)-Metiltransferases/efeitos dos fármacos , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA , Metilases de Modificação do DNA/efeitos dos fármacos , Metilases de Modificação do DNA/metabolismo , Enzimas Reparadoras do DNA/efeitos dos fármacos , Enzimas Reparadoras do DNA/metabolismo , Expressão Gênica , Histonas/genética , Humanos , Imunoprecipitação , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição Gênica , Proteínas Supressoras de Tumor/efeitos dos fármacos , Proteínas Supressoras de Tumor/metabolismo
19.
Clin Adv Hematol Oncol ; 5(7): 544-52, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17679928

RESUMO

Aberrant DNA methylation is one of the molecular hallmarks of cancer and leukemia. By repressing gene expression, it is considered a functional equivalent to the physical inactivation of tumor suppressor genes by deletions or mutations. To clinically exploit this process, compounds with DNA hypomethylating properties have been evaluated both in the laboratory and the clinic. Two such agents, 5-azacytidine and 5-aza-2'-deoxycytidine, are currently approved by the US Food and Drug Administration for the treatment of patients with myelodysplastic syndromes. Ongoing studies are evaluating alternative dosing schedules for these drugs and the activity and safety of this class of agent in combination with histone deacetylase inhibitors. Here we summarize the experience of hypomethylating agents in myelodysplastic syndromes.


Assuntos
Antimetabólitos Antineoplásicos/farmacologia , Metilação de DNA/efeitos dos fármacos , Metilases de Modificação do DNA/efeitos dos fármacos , Síndromes Mielodisplásicas/tratamento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Azacitidina/análogos & derivados , Azacitidina/farmacologia , Decitabina , Inibidores de Histona Desacetilases , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto
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