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1.
Expert Opin Ther Pat ; 29(8): 623-641, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31353978

RESUMO

Introduction: About 20 patents have been published from 2013 to 2018 for developing advanced cancer therapeutics by targeting tubulin polymerization. Currently, there are several tubulin inhibitors that are in the drug development pipeline for various cancers alone or in combination including antibody-conjugated drugs (ACDs). Areas covered: Important patents focusing on the development of tubulin inhibitors published from 2013 to 2018 are covered. This review mainly focuses on the tubulin inhibitors that are being synthesized and studied in cancer research along with their structures and their phases of development in preclinical and clinical research. Expert opinion: Regulation of microtubules is important for cell division, cell motility, intracellular transport, and cell shape maintenance. Modulating its activity proved to be very effective in various diseases including different types of cancers. Microtubules are composed of two units, namely, alpha and beta-tubulin, and modifications at these ends affect both its functions and dynamics. A number of compounds that have been designed and synthesized bearing various heterocyclic scaffolds have been proven to modulate its activity and have emerged as potent tubulin inhibitors. This encourages more to study microtubules in order to find a variety of novel, potent compounds as anticancer drugs.


Assuntos
Antineoplásicos/farmacologia , Neoplasias/tratamento farmacológico , Moduladores de Tubulina/farmacologia , Animais , Antineoplásicos/química , Desenho de Drogas , Desenvolvimento de Medicamentos/métodos , Humanos , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Neoplasias/patologia , Patentes como Assunto , Relação Estrutura-Atividade , Tubulina (Proteína)/efeitos dos fármacos , Tubulina (Proteína)/metabolismo , Moduladores de Tubulina/química
2.
BMC Complement Altern Med ; 19(1): 116, 2019 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-31164129

RESUMO

BACKGROUND: Allium species are magnificently nutritious and are commonly used as a part of the diet in Iran. They have health enhancing benefits including anticancer properties due to the presence of numerous bioactive compounds. Herein, we investigated in vitro and in vivo anticancer properties of Allium bakhtiaricum extracts. METHODS: Anti-growth activity of different fractions was explored in vitro on different cancerous cells using MTT assay, Annexin V/PI and SA-ß-gal staining, Western blotting, flowcytometric and immunofluorescence microscopic evaluations. In vivo antitumor activity was investigated in BALB/c mice bearing 4 T1 mammary carcinoma cells. RESULTS: We demonstrated that chloroformic and ethyl acetate fractions exert cytotoxic activity toward MDA-MB-231 cells, the most sensitive cell line, after 72 h of treatment with IC50 values of 0.005 and 0.006 mg/ml, respectively. Incubation of MDA-MB-231 cells with » and ½ IC50-72h concentrations of each fraction resulted in a significant G2/M cell cycle arrest. » IC50-72h concentration of the chloroform fraction led to the disruption of polymerization in mitotic microtubules. Exposure of human breast cancer cells to different concentrations of the extracts at different incubation times did not induce apoptosis, autophagy or senescence. Our in vivo study revealed that administration of the chloroform extract at a dose of 1 mg/kg/day strongly suppressed mammary tumor progression and decreased the number of proliferative cells in the lung tissues indicating its anti-metastatic effect. CONCLUSION: Our findings imply that the chloroform fraction of Allium bakhtiaricum possesses the suppressive action on breast cancer through mitotic cell cycle arrest suggesting a mechanism associated with disturbing microtubule polymerization.


Assuntos
Allium/química , Antineoplásicos Fitogênicos/análise , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Microtúbulos/efeitos dos fármacos , Metástase Neoplásica , Extratos Vegetais/química , Extratos Vegetais/farmacologia
3.
Expert Opin Investig Drugs ; 28(6): 513-523, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31159588

RESUMO

INTRODUCTION: The management of non-small cell lung cancer (NSCLC) has been substantially improved in the last few years; it has been revolutionized by a patient-tailored approach, especially in the oncogene addicted disease, and by novel combinations containing immune checkpoint inhibitors. However, chemotherapy still represents a mainstay that persists over the decades with limited novelties. Tubulin inhibitors belong to different sub-classes of drugs that share the capability to interfere with mitosis by a direct action on the microtubule system. Among them, taxanes and vinca alkaloids still have a prominent role in clinical practice. AREAS COVERED: This review summarizes the mechanisms of action, current role and future directions of microtubule targeting agents; we focus on investigational agents in phase I and II clinical trials. EXPERT OPINION: Chemotherapy maintains a pivotal role in the treatment of NSCLC. New generation agents that have the potential to overcome the mechanisms of resistance to the available drugs may provide new therapeutic opportunities. Predictive biomarkers derived from combination strategies and phase III studies are necessary going forward.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Moduladores de Tubulina/administração & dosagem , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/farmacologia , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Desenho de Drogas , Drogas em Investigação/administração & dosagem , Drogas em Investigação/farmacologia , Humanos , Neoplasias Pulmonares/patologia , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Moduladores de Tubulina/farmacologia
4.
Top Curr Chem (Cham) ; 377(3): 15, 2019 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-31073777

RESUMO

As the emergence of resistance to clinical cancer treatments poses a significant problem in cancer management, there is a constant need to explore novel anticancer agents which have the ability to overcome multidrug resistance (MDR) mechanisms. The search for the development of novel isatin-based antitumor agents accelerated after the approval by the Food and Drug Administration (FDA) of sunitinib malate, a C-3 isatin derivative, as a multitargeted receptor tyrosine kinase inhibitor. However, it is interesting to note that, over the last decade, various N-substituted analogs of isatin with intact carbonyl functionalities have been found to show more promising anticancer potential than its C-3 derivatives. Microtubule-targeting agents are a class of anticancer drugs which affect mitosis by targeting microtubules and suppressing their dynamic behavior. This review presents a systematic compilation of the in vitro cytotoxic and anticancer properties of various N-substituted isatins and illustrates their mechanism of action to overcome MDR by acting as microtubule-destabilizing agents. Predictions of the biological activities and cytotoxic effects of potential N-substituted isatins against various cancer cell lines have also been performed using the PASS computer-aided drug discovery program. Findings from such in vitro and in silico studies will act as a guide for the development of structure-activity relationship and will facilitate the design and exploration of more potent analogs of isatin with high potency and lower side effects for treatment of drug-resistant cancer. Mechanism of action of N-substituted isatin as microtubule-destabilizing agent on tumor cells. N-Substituted isatins bind to colchicine binding site on ß-tubulin, which inhibits microtubule polymerization and thereby destabilizes microtubule dynamics, resulting in mitotic arrest leading to tumor cell growth suppression.


Assuntos
Antineoplásicos/farmacologia , Simulação por Computador , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Isatina/farmacologia , Relação Quantitativa Estrutura-Atividade , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Técnicas In Vitro , Isatina/química , Microtúbulos/efeitos dos fármacos
5.
Int J Mol Sci ; 20(8)2019 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-31010006

RESUMO

Vincristine is used in the clinical treatment of colon cancer, especially in patients diagnosed in the advanced phase of cancer development. Unfortunately, similar to other agents used during antitumor therapy, vincristine might induce chemoresistance. Studies of this process focus mainly on the analysis of the molecular mechanisms within cancer, usually ignoring the role of stromal cells. Our present findings confirm that vincristine stimulates the secretion of tumor growth factors class beta and interleukin-6 from cancer-associated fibroblasts as a result of paracrine stimulation by cancer cells. Based on alterations in morphology, modulation of capillary formation, and changes in endothelial and mesenchymal marker profile, our findings demonstrate that higher levels of tumor growth factor-ßs and interleukin-6 enhance cancer-associated fibroblast-like cell formation through endothelial-mesenchymal transition and that nonsteroidal anti-inflammatory drug treatment (aspirin and ibuprofen) is able to inhibit this phenomenon. The process appears to be regulated by the rate of microtubule polymerization, depending on ß-tubulin composition. While higher levels of tubulin-ß2 and tubulin-ß4 caused slowed polymerization and reduced the level of factors secreted to the extracellular matrix, tubulin-ß3 induced the opposite effect. We conclude that nonsteroidal anti-inflammatory drugs should be considered for use during vincristine monotherapy in the treatment of patients diagnosed with colorectal cancer.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Fibroblastos Associados a Câncer/patologia , Vincristina/farmacologia , Fibroblastos Associados a Câncer/efeitos dos fármacos , Fibroblastos Associados a Câncer/metabolismo , Comunicação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Transdiferenciação Celular/efeitos dos fármacos , Neoplasias do Colo/patologia , Meios de Cultivo Condicionados/farmacologia , Citocinas/metabolismo , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Endotélio/efeitos dos fármacos , Endotélio/patologia , Humanos , Mesoderma/efeitos dos fármacos , Mesoderma/patologia , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Polimerização , Tubulina (Proteína)/metabolismo
6.
Eur J Med Chem ; 173: 1-14, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30981112

RESUMO

Further optimization of the trimethoxyphenyl scaffold of parent chalcone compound (2a) by introducing a pyridine ring afforded a series of novel pyridine-chalcone derivatives as potential anti-tubulin agents. All the target compounds were evaluated for their antiproliferative activities. Among them, representative compound 16f exhibited the most potent activity with the IC50 values ranging from 0.023 to 0.045 µM against a panel of cancer cell lines. Further mechanism study results demonstrated that compound 16f effectively inhibited the microtubule polymerization by binding to the colchicine site of tubulin. Moreover, cellular mechanism studies disclosed that 16f caused G2/M phase arrest, induced cell apoptosis and disrupted the intracellular microtubule network. Also, 16f reduced the cell migration and disrupted the capillary-like tube formation of human umbilical vein endothelial cells (HUVECs). Importantly, 16f significantly inhibited tumor growth in H22 xenograft models without apparent toxicity, which was stronger than the reference compound CA-4, indicating that it is worthy to investigate 16f as a potent microtubule-destabilizing agent for cancer therapy.


Assuntos
Antineoplásicos/farmacologia , Chalcona/farmacologia , Desenho de Drogas , Microtúbulos/efeitos dos fármacos , Piridinas/farmacologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Chalcona/síntese química , Chalcona/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Células K562 , Masculino , Camundongos , Camundongos Endogâmicos ICR , Microtúbulos/metabolismo , Modelos Moleculares , Estrutura Molecular , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Piridinas/síntese química , Piridinas/química , Relação Estrutura-Atividade , Cicatrização/efeitos dos fármacos
7.
Methods Mol Biol ; 1957: 271-289, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30919360

RESUMO

Primary cilia (PC) are microtubule-based organelles that behave like a cellular antenna controlling key signaling pathways during development and tissue homeostasis. The ciliary membrane is highly enriched for G protein-coupled receptors (GPCRs), and PC are a crucial signaling compartment for this large receptor family. Downstream effectors of GPCR signaling are also present in cilia, and evidence obtained by our labs and others demonstrated that ß-arrestin (ßarr) family members are differentially recruited to PC and have investigated the role of GPCR activation in this process. In this chapter, we provide methods based on fluorescence microscopy on fixed or live cells suitable for investigating targeting and recruitment of ßarrs at PC.


Assuntos
Corpos Basais/metabolismo , Centrossomo/metabolismo , Cílios/metabolismo , Microscopia de Fluorescência/métodos , beta-Arrestina 2/metabolismo , Animais , Corpos Basais/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Centrossomo/efeitos dos fármacos , Cílios/efeitos dos fármacos , DNA/metabolismo , Recuperação de Fluorescência Após Fotodegradação , Proteínas de Fluorescência Verde/metabolismo , Hipocampo/citologia , Humanos , Camundongos , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Plasmídeos/metabolismo , Somatostatina/farmacologia
8.
Planta ; 249(5): 1551-1563, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30729290

RESUMO

MAIN CONCLUSION: A novel method for culturing ovules of Gossypium barbadense allowed in vitro comparisons with Gossypium hirsutum and revealed variable roles of microtubules in controlling cotton fiber cell expansion. Cotton fibers undergo extensive elongation and secondary wall thickening as they develop into our most important renewable textile material. These single cells elongate at the apex as well as elongating and expanding in diameter behind the apex. These multiple growth modes represent an interesting difference compared to classical tip-growing cells that needs to be explored further. In vitro ovule culture enables experimental analysis of the controls of cotton fiber development in commonly grown Gossypium hirsutum cotton, but, previously, there was no equivalent system for G. barbadense, which produces higher quality cotton fiber. Here, we describe: (a) how to culture the ovules of G. barbadense successfully, and (b) the results of an in vitro experiment comparing the role of microtubules in controlling cell expansion in different zones near the apex of three types of cotton fiber tips. Adding the common herbicide fluridone, 1-Methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone, to the medium supported G. barbadense ovule culture, with positive impacts on the number of useful ovules and fiber length. The effect is potentially mediated through inhibited synthesis of abscisic acid, which antagonized the positive effects of fluridone. Fiber development was perturbed by adding colchicine, a microtubule antagonist, to ovules of G. barbadense and G. hirsutum cultured 2 days after flowering. The results supported the zonal control of cell expansion in one type of G. hirsutum fiber tip and highlighted differences in the role of microtubules in modulating cell expansion between three types of cotton fiber tips.


Assuntos
Gossypium/citologia , Gossypium/metabolismo , Microtúbulos/metabolismo , Fibra de Algodão , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Gossypium/efeitos dos fármacos , Microtúbulos/efeitos dos fármacos , Piridonas/farmacologia
9.
Oncogene ; 38(22): 4352-4365, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30770899

RESUMO

Anti-microtubule agents are frequently used as anticancer therapeutics. Cell death induced by these agents is considered to be due to sustained mitotic arrest caused by the activation of spindle assembly checkpoint (SAC). However, some cell types are resistant to mitotic cell death. Cells' ability to escape mitotic arrest (mitotic slippage) is thought to be a major mechanism contributing to this resistance. Here, we show that resistance to cell death induced by anti-mitotic agents is not linked to cells' capacity to undergo mitotic slippage as generally believed but is dependent on the state of BimEL regulation during mitosis. While transcriptional repression of BimEL in the mitotic death-resistant cells involves polycomb repressive complex 2 (PRC2)-mediated histone trimethylation, the BimEL protein is destabilized by cullin 1/4A-ßTrCP-dependent degradation involving activation of cullin 1/4A by neddylation. These results imply that pharmacological augmentation of BimEL activity in anti-microtubule drug-resistant tumors may have important therapeutic implications.


Assuntos
Proteína 11 Semelhante a Bcl-2/genética , Morte Celular/genética , Resistência a Medicamentos/genética , Microtúbulos/genética , Células A549 , Antineoplásicos/farmacologia , Proteínas de Ciclo Celular/genética , Morte Celular/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Resistência a Medicamentos/efeitos dos fármacos , Células HEK293 , Células HeLa , Histonas/genética , Humanos , Pontos de Checagem da Fase M do Ciclo Celular/genética , Metilação/efeitos dos fármacos , Microtúbulos/efeitos dos fármacos , Mitose/efeitos dos fármacos , Mitose/genética , Complexo Repressor Polycomb 2/genética , Fuso Acromático/efeitos dos fármacos , Fuso Acromático/genética , Transcrição Genética/efeitos dos fármacos , Transcrição Genética/genética
10.
Biol Pharm Bull ; 42(5): 814-818, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30787205

RESUMO

In addition to their major targets, clinically effective drugs may have unknown off-targets. By identifying such off-targets it may be possible to repurpose approved drugs for new indications. We are interested in the Golgi apparatus as a novel target for cancer therapy, but there is a paucity of candidate Golgi-disrupting drugs. Here, we aimed to identify Golgi-disrupting compounds from a panel of 34 approved anticancer drugs by using HBC-4 human breast cancer cells and immunofluorescence microscopy to visualize the Golgi apparatus. The screen identified five drugs having Golgi-disrupting activity. Four of them were vinca alkaloids (vinorelbine, vindesine, vincristine and vinblastine), and the fifth drug was eribulin. This is the first study to demonstrate that vinorelbine, vindesine and eribulin possess Golgi-disrupting activity. The 5 drugs are known to inhibit tubulin polymerization and to induce microtubule depolymerization. Interestingly, a microtubule-stabilizer paclitaxel did not induce Golgi-disruption, suggesting that the three-dimensionally preserved microtubules are partly responsible for maintaining the Golgi complex. Concerning eribulin, a noteworthy drug because of its high clinical efficacy against advanced breast cancer, we further confirmed its Golgi-disrupting activity in 3 different human breast cancer cell lines, BSY-1, MDA-MB-231 and MCF-7. Golgi-disruption may contribute to anticancer efficacy of eribulin. In conclusion, the present study revealed that 4 vinca alkaloids and eribulin possessed potential Golgi-disrupting activity among a panel of 34 approved anticancer drugs. Other drugs covering various molecular-targeted drugs and classical DNA-damaging drugs showed no Golgi-disrupting effect. These results suggest that tubulin polymerization-inhibitors might be promising candidate drugs with Golgi-disrupting activity.


Assuntos
Antineoplásicos/farmacologia , Complexo de Golgi/efeitos dos fármacos , Moduladores de Tubulina/farmacologia , Antineoplásicos/efeitos adversos , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Furanos/farmacologia , Complexo de Golgi/metabolismo , Ensaios de Triagem em Larga Escala , Humanos , Cetonas/farmacologia , Células MCF-7 , Microtúbulos/efeitos dos fármacos , Paclitaxel/farmacologia , Vimblastina/farmacologia , Vincristina/farmacologia , Vindesina/farmacologia , Vinorelbina/farmacologia
11.
Biomed Pharmacother ; 112: 108645, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30798125

RESUMO

Glioblastoma is the most common and the most malignant form of brain tumor. This devastating tumor results in death within a year after diagnosis. Although the tumor mass can be surgically removed, glioma cells invade other areas in the brain leading to tumor recurrence and poor prognosis. Therefore, new agents that can overcome cancer cell invasion are urgently required. Phyllanthus taxodiifolius Beille (P. taxodiifolius), has been reported to have potent anti-cancer activities. However, its effects on glioblastoma cells and its underlying mechanisms have never been revealed. Here we investigated the effect and underlying mechanisms of P. taxodiifolius extract on aggressive properties of the glioblastoma, including adhesion, migration, and invasion. P. taxodiifolius extract disrupted adhesion, delayed migration and interfered with the invasion of glioblastoma cells. In addition, the extract suppressed microtubule dynamics as shown by live imaging of a microtubule plus tip protein and decreased focal adhesion by decreasing focal adhesion kinase activity. Our study is the first evidence showing that P. taxodiifolius extract suppresses invasive properties of glioblastoma cells by disrupting microtubule structure and interfering with microtubule dynamics, suggesting the possibility to further develop P. taxodiifolius and its bioactive compounds as an anti-cancer drug targeting microtubules in glioblastoma.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Encefálicas/patologia , Glioblastoma/patologia , Microtúbulos/efeitos dos fármacos , Phyllanthus/química , Componentes Aéreos da Planta/química , Extratos Vegetais/farmacologia , Animais , Antineoplásicos Fitogênicos/isolamento & purificação , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Microscopia Confocal , Microtúbulos/ultraestrutura , Invasividade Neoplásica , Extratos Vegetais/isolamento & purificação , Ratos
12.
Proc Natl Acad Sci U S A ; 116(9): 3774-3783, 2019 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-30808763

RESUMO

Establishing causal links between bacterial metabolites and human intestinal disease is a significant challenge. This study reveals the molecular basis of antibiotic-associated hemorrhagic colitis (AAHC) caused by intestinal resident Klebsiella oxytoca Colitogenic strains produce the nonribosomal peptides tilivalline and tilimycin. Here, we verify that these enterotoxins are present in the human intestine during active colitis and determine their concentrations in a murine disease model. Although both toxins share a pyrrolobenzodiazepine structure, they have distinct molecular targets. Tilimycin acts as a genotoxin. Its interaction with DNA activates damage repair mechanisms in cultured cells and causes DNA strand breakage and an increased lesion burden in cecal enterocytes of colonized mice. In contrast, tilivalline binds tubulin and stabilizes microtubules leading to mitotic arrest. To our knowledge, this activity is unique for microbiota-derived metabolites of the human intestine. The capacity of both toxins to induce apoptosis in intestinal epithelial cells-a hallmark feature of AAHC-by independent modes of action, strengthens our proposal that these metabolites act collectively in the pathogenicity of colitis.


Assuntos
Enterocolite Pseudomembranosa/genética , Enterotoxinas/metabolismo , Interações entre Hospedeiro e Microrganismos/genética , Klebsiella oxytoca/genética , Animais , Benzodiazepinonas/metabolismo , Benzodiazepinonas/toxicidade , Dano ao DNA/efeitos dos fármacos , Enterocolite Pseudomembranosa/microbiologia , Enterocolite Pseudomembranosa/patologia , Enterotoxinas/biossíntese , Células Epiteliais/microbiologia , Células Epiteliais/patologia , Humanos , Intestinos/microbiologia , Intestinos/patologia , Infecções por Klebsiella/genética , Infecções por Klebsiella/microbiologia , Klebsiella oxytoca/metabolismo , Klebsiella oxytoca/patogenicidade , Camundongos , Microtúbulos/efeitos dos fármacos , Oxiquinolina/análogos & derivados , Oxiquinolina/metabolismo , Oxiquinolina/toxicidade , Peptídeos/metabolismo , Peptídeos/toxicidade
13.
Eur J Pharmacol ; 846: 86-99, 2019 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-30641059

RESUMO

Many ent-kaurane diterpenoids exhibit notable antitumor activity in vitro and in vivo, and some have been used as cancer therapeutic agents in China. In this study, we identified a novel molecular target of leukamenin E, an ent-kaurane diterpenoid, using an available whole-cell model in combination with immunofluorescence imaging and mass spectrometry (MS). The cytoskeleton-disrupting drugs cytochalasin B and colchicine caused the depolymerization of microfilaments and the collapse of microtubules and vimentin filaments, respectively, but had little effects on HepG2 and NCI-H1299 cells spreading as well as keratin filament (KF) reassembly, indicating that KFs are involved in cell spreading. Leukamenin E blocked HepG2 and NCI-H1299 cells adhesion/spreading and KF reassembly at subtoxic concentrations, indicating that leukamenin E may target KFs. Moreover, leukamenin E, at 3 µM for 24 h or 10 µM for 3 h, induced massive KF depolymerization in well-spread HepG2 and NCI-H1299 cells treated with/without cytochalasin B and colchicine. MS analysis indicated that leukamenin E could covalently modify amino acid residue(s) in a synthetic peptide based on keratin 1 and keratin 10 sequences, suggesting that covalent modification of the synthetic peptide by leukamenin E caused assembly inhibition or disrupted KF polymerization in HepG2 and NCI-H1299 cells. In addition, acridine orange/ethidium bromide staining and western blotting confirmed that there was no correlation between the KF-disrupting effects and apoptosis or keratin expression. Thus, we propose that leukamenin E is a novel inhibitor of KF assembly, and as such, can serve as a chemical probe of KF functions and a potential molecular target for ent-kaurane diterpenoid-based therapeutics.


Assuntos
Citoesqueleto de Actina/metabolismo , Adesão Celular/efeitos dos fármacos , Diterpenos de Caurano/farmacologia , Queratinas/metabolismo , Fatores de Despolimerização de Actina , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Colchicina/farmacologia , Citocalasina B/farmacologia , Células Hep G2 , Humanos , Espectrometria de Massas/métodos , Microscopia de Fluorescência/métodos , Microtúbulos/efeitos dos fármacos , Vimentina/metabolismo
14.
Environ Toxicol ; 34(4): 469-475, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30614199

RESUMO

Cadmium (Cd) is considered a possible etiological factor in neurodegenerative diseases. However, the exact mechanism by which Cd induces neurotoxicity is not well elucidated. In this study, Neuro-2a cells were treated with 0, 10, 20, and 40 µM cadmium chloride for 24 hours to investigate the effects of Cd on the cytoskeleton of nerve cells. MTT assay and ELISA assay were used to examine cell viability and release of lactate dehydrogenase (LDH) from cells, respectively. Results showed that Cd reduced cell viability and increased the release of LDH in a dose-dependent manner (P < 0.05). The morphology of treated cell was damaged as indicated by cell collapse and dimensionality reduction. Moreover, the axonal spines and normal features of Cd-treated neurons disappeared. We checked the ultrastructure of Neuro-2a cells and found that Cd-induced swelling, membrane damage, overflow of cytoplasm contents, and cell fragmentation. Damaged mitochondria, expanded endoplasmic reticulum, and abnormal microfilaments were found in Cd-treated cells rather than in untreated cells. Compared with the control group, the relative release of glutamate in the supernatant after Cd treatment was reduced, indicating that Cd exposure could reduce the release of glutamate by inhibiting the function of nerve-2a cells. Cd decreased the mRNA and protein expression levels of cytoskeletal proteins including DBN, SYP, and TAU, which might promote cytoskeleton alterations in Cd-treated cells. In conclusion, Cd-induced actin cytoskeleton alterations and dysfunction of cultured neurons. The results of the present study provide new insights for the investigation of Cd-induced neurotoxicity.


Assuntos
Citoesqueleto de Actina/efeitos dos fármacos , Cádmio/toxicidade , Poluentes Ambientais/toxicidade , Neurônios/efeitos dos fármacos , Citoesqueleto de Actina/ultraestrutura , Animais , Axônios/efeitos dos fármacos , Axônios/ultraestrutura , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Camundongos , Microtúbulos/efeitos dos fármacos , Microtúbulos/ultraestrutura , Neurônios/ultraestrutura , Síndromes Neurotóxicas/patologia
15.
Cell Mol Life Sci ; 76(6): 1169-1183, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30599068

RESUMO

Senescent cells undergo structural and functional changes that affect essentially every aspect of cell physiology. To date, the impact of senescence on the cytoskeleton is poorly understood. This study evaluated the cytoskeleton in two independent cellular models of kidney epithelium senescence. Our work identified multiple senescence-related alterations that impact microtubules and filamentous actin during interphase. Both filamentous systems reorganized profoundly when cells became senescent. As such, microtubule stability increased during senescence, making these filaments more resistant to disassembly in the cold or by nocodazole. Microtubule stabilization was accompanied by enhanced α-tubulin acetylation on lysine 40 and the depletion of HDAC6, the major deacetylase for α-tubulin lysine 40. Rho-associated kinase Rock1 is an upstream regulator that modulates key properties of the cytoplasmic cytoskeleton. Our research shows that Rock1 concentrations were reduced significantly in senescent cells, and we revealed a mechanistic link between microtubule stabilization and Rock1 depletion. Thus, Rock1 overexpression partially restored the cold sensitivity of microtubules in cells undergoing senescence. Additional components relevant to microtubules were affected by senescence. Specifically, we uncovered the senescence-related loss of the microtubule nucleating protein γ-tubulin and aberrant formation of γ-tubulin foci. Concomitant with the alterations of microtubule and actin filaments, senescent cells displayed functional changes. In particular, cell migration was impaired significantly in senescent cells. Taken together, our study identified new senescence-associated deficiencies of the microtubule and actin cytoskeleton, provided insights into the underlying molecular mechanisms and demonstrated functional consequences that are important to the physiology and function of renal epithelial cells.


Assuntos
Citoesqueleto de Actina/metabolismo , Senescência Celular , Microtúbulos/metabolismo , Tubulina (Proteína)/metabolismo , Citoesqueleto de Actina/efeitos dos fármacos , Animais , Linhagem Celular , Células Epiteliais/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Túbulos Renais Proximais/citologia , Microscopia Confocal , Microtúbulos/efeitos dos fármacos , Nocodazol/farmacologia , Suínos , Moduladores de Tubulina/farmacologia , Quinases Associadas a rho/genética , Quinases Associadas a rho/metabolismo
16.
Org Biomol Chem ; 17(6): 1519-1530, 2019 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-30681116

RESUMO

We report the identification of 14 novel anticancer agents through established computational anticancer cell-based models. Among these novel hits, the compound G03 exhibits stronger inhibitory effects on the proliferation of MCF-7, HepG2, MDA-MB-231, HCTT116, and HeLa as compared with the FDA-approved sorafenib, with IC50 values of 4.61, 3.20, 2.82, 2.98, and 2.90 µM, respectively. The tubulin protein was validated to be a target of G03 using SPR, tubulin polymerization, immunofluorescence, and western blot assays. G03 is a novel structurally simple anticancer agent with unusual microtubule-stabilizing effects. Our study demonstrated the identification of bioactive small molecules by computational phenotypic modeling, which represents a feasible route toward innovative leads for chemical biology and medicinal chemistry.


Assuntos
Antineoplásicos/farmacologia , Bioensaio , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Simulação de Acoplamento Molecular , Antineoplásicos/metabolismo , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Conformação Proteica , Tubulina (Proteína)/química , Tubulina (Proteína)/metabolismo , Interface Usuário-Computador
17.
Adv Colloid Interface Sci ; 263: 95-130, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30530177

RESUMO

Cancer is the second cause of death worldwide, exceeded only by cardiovascular diseases. The prevalent treatment currently used against metastatic cancer is chemotherapy. Among the most studied drugs that inhibit neoplastic cells from acquiring unlimited replicative ability (a hallmark of cancer) are the taxanes. They operate via a unique molecular mechanism affecting mitosis. In this review, we show this mechanism for one of them, paclitaxel, and for other (non-taxanes) anti-mitotic drugs. However, the use of paclitaxel is seriously limited (its bioavailability is <10%) due to several long-standing challenges: its poor water solubility (0.3 µg/mL), its being a substrate for the efflux multidrug transporter P-gp, and, in the case of oral delivery, its first-pass metabolism by certain enzymes. Adequate delivery methods are therefore required to enhance the anti-tumor activity of paclitaxel. Thus, we have also reviewed drug delivery strategies in light of the various physical, chemical, and enzymatic obstacles facing the (especially oral) delivery of drugs in general and paclitaxel in particular. Among the powerful and versatile platforms that have been developed and achieved unprecedented opportunities as drug carriers, microemulsions might have great potential for this aim. This is due to properties such as thermodynamic stability (leading to long shelf-life), increased drug solubilization, and ease of preparation and administration. In this review, we define microemulsions and nanoemulsions, analyze their pertinent properties, and review the results of several drug delivery carriers based on these systems.


Assuntos
Antineoplásicos Fitogênicos/farmacocinética , Sistemas de Liberação de Medicamentos/métodos , Mitose/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Paclitaxel/farmacocinética , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Disponibilidade Biológica , Transporte Biológico , Estabilidade de Medicamentos , Emulsões , Glicerol/administração & dosagem , Glicerol/efeitos adversos , Glicerol/análogos & derivados , Humanos , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Microtúbulos/ultraestrutura , Nanoestruturas/administração & dosagem , Nanoestruturas/química , Neoplasias/metabolismo , Neoplasias/patologia , Paclitaxel/química , Paclitaxel/farmacologia , Solubilidade , Solventes/administração & dosagem , Solventes/efeitos adversos , Células Tumorais Cultivadas
18.
Biochem Biophys Res Commun ; 508(3): 986-990, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30545630

RESUMO

Surgery and chemotherapy are the gold-standard treatments for ovarian cancer. The major cause of treatment failure in patients with ovarian cancer is tumoral heterogeneity and drug resistance. Paclitaxel (PTX) is one of the most commonly used first-line drugs for ovarian cancer chemotherapy. Unfortunately, the mechanisms of PTX chemoresistance remain unclear. Here, we examined the effects of post-translational neddylation on the sensitivity of ovarian cancer cells (OCCs) to PTX-induced apoptosis. Disruption of protein neddylation with the first-in-class inhibitor MLN4924 dramatically neutralized PTX-mediated antiproliferative, antimigration, and apoptotic effects in human OCCs. Moreover, MLN4924 treatment interrupted PTX-induced microtubule polymerization. Importantly, two neddylation conjugating E2 enzymes, UBE2M and UBE2F, were found to play essential roles in PTX-induced cytotoxicity and tubulin polymerization in OCCs. In summary, our findings demonstrated that disruption of protein neddylation by MLN4924 conferred resistance to PTX and provided insights into the potential mechanisms of PTX chemoresistance in ovarian cancer.


Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Ciclopentanos/farmacologia , Microtúbulos/efeitos dos fármacos , Proteína NEDD8/metabolismo , Neoplasias Ovarianas/tratamento farmacológico , Paclitaxel/uso terapêutico , Pirimidinas/farmacologia , Apoptose , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos , Feminino , Humanos , Microtúbulos/metabolismo , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Polimerização/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Enzimas de Conjugação de Ubiquitina/antagonistas & inibidores
20.
Int J Nanomedicine ; 14: 17-32, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30587982

RESUMO

Background: Docetaxel (DOC) is widely used as a chemotherapy drug for various tumors, including gastric cancer (GC), but the clinical application of DOC has been limited due to the hydrophobicity of the drug. We aimed to formulate a multifunctional nanoparticle (NP) system to reduce the side effects of the chemotherapy agent, to promote synergistic therapeutic effects, and to achieve targeted delivery of the therapy. Methods: The polyethylene glycol-poly(ε-caprolactone) NPs (PEG-PCL NPs) were prepared by a ring opening copolymerization technique and were then conjugated with a programmed death-ligand 1 (PD-L1) monoclonal antibody (mAb). The effects of the surface coating on particle size, size distribution, zeta potential, drug encapsulation efficiency, loading capacity, and the drug release kinetics were investigated. By using a panel of PD-L1-expressing human GC cell lines and PD-L1-overexpressing cells, we studied cellular uptake, cytotoxic effects, and cellular apoptosis in the presence of PD-L1 mAb-conjugated NPs. Results: The characterization of the structure and biological functions of DOC-PEG-PCL-mAb NPs was investigated in vitro. X-ray photoelectron spectroscopy validated the presence of the PD-L1 mAbs on the NP surface. The cellular uptake analysis showed that the antibody-conjugated NPs achieved significantly higher cellular uptake. The results of an in vitro cytotoxicity experiment on three GC lines further proved the targeting effects of the antibody conjugation. In addition, we found that the DOC-PEG-PCL-mAb NPs induced cell apoptosis and enhanced G2-M arrest in cancer cells, indicating the inhibition of microtubule synthesis. When compared with the control groups, DOC-PEG-PCL-mAb NPs are more effective in inhibiting PD-L1 expression in GC cells. Conclusion: Our results reported here highlight the biological and clinical potential of DOC-PEG-PCL-mAb NPs using PD-L1 mAbs in GC treatment.


Assuntos
Anticorpos Monoclonais/metabolismo , Antineoplásicos/uso terapêutico , Antígeno B7-H1/metabolismo , Sistemas de Liberação de Medicamentos , Nanopartículas/química , Neoplasias Gástricas/tratamento farmacológico , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Docetaxel/farmacologia , Docetaxel/uso terapêutico , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Humanos , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Nanopartículas/ultraestrutura , Tamanho da Partícula , Espectroscopia Fotoeletrônica , Poliésteres/química , Polietilenoglicóis/química , Resultado do Tratamento
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