RESUMO
The discovery and development of effective novel compounds is paramount in oncology for improving cancer therapy. In this study, we developed a new derivative of spiroindolone (7',8'-Dimethoxy-1',3'-dimethyl-1,2,3',4'-tetrahydrospiro[indole-3,5'- pyrazolo[3,4-c]isoquinolin]-2-one) and evaluated its anticancer- and immunomodulatory potential in a vitro model of chronic leukemia. We utilized the chronic leukemia cell line K562, as well as non-cancerous peripheral blood mononuclear cells (PBMC) and Vero cells (kidney epithelium of Cercopithecus aethiops). We assessed the cytotoxicity of the compound using the MTT assay, and performed cell cycle assays to determine its impact on different stages of the cell cycle. To evaluate its antineoplastic activity, we conducted a colony formation test to measure the effect of the compound on the clonal growth of cancer cells. Furthermore, we evaluated the immunomodulatory activity of the compound by measuring the levels of pro and anti-inflammatory cytokines. The study findings demonstrate that the spiroindolone-derived compound exerted noteworthy cytotoxic effects against K562 cells, with an IC50 value of 25.27 µg/mL. Additionally, it was observed that the compound inhibited the clonal proliferation of K562 cells while displaying minimal toxicity to normal cells. The compound exhibited its antiproliferative activity by inducing G2/M cell cycle arrest, preventing the entry of K562 cells into mitosis. Notably, the compound demonstrated an immunomodulatory effect by upregulating the production of cytokines IL-6 and IL-12/23p40. In conclusion, the spiroindolone-derived compound evaluated in this study has demonstrated significant potential as a therapeutic agent for the treatment of chronic myeloid leukemia. Further investigations are warranted to explore its clinical applications.
Assuntos
Antineoplásicos , Leucemia Mielogênica Crônica BCR-ABL Positiva , Humanos , Animais , Chlorocebus aethiops , Leucócitos Mononucleares , Células Vero , Proliferação de Células , Apoptose , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Células K562 , Citocinas/farmacologia , Indóis/farmacologiaRESUMO
Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) infections are widely distributed throughout the world. EBV is linked to various hematological and autoimmune disorders whereas CMV might play important role in the progression of chronic hematological diseases, such as hemoglobinopathies, lymphomas, myelomas, hemophilia, and aplastic and sickle cell anemia. Both viruses produce a viral homolog of human interleukin-10 that can cause general suppression of immune response, increasing susceptibility to other infections. These viruses can remain latent in the host cells and be reactivated when the host immune system is compromised. Studies showing the impact of CMV and EBV infections on hematological disorders are scarce and unclear in the context of coinfection. This review intends to present the biology, prevalence, and impact of CMV and EBV infections in patients with hematological diseases.
Assuntos
Infecções por Citomegalovirus/complicações , Infecções por Citomegalovirus/epidemiologia , Infecções por Vírus Epstein-Barr/complicações , Infecções por Vírus Epstein-Barr/epidemiologia , Doenças Hematológicas/virologia , Humanos , Modelos Biológicos , PrevalênciaRESUMO
ANKHD1 is highly expressed in various cancers such as leukemia and multiple myeloma. Silencing of ANKHD1 expression leads to decreased cell proliferation and accumulation of cells at the S phase. In this study we found ANKHD1 expression to be higher at the S phase, suggesting it to be an S phase protein. We observed that ANKHD1 interacts with histone promoter regions and its inhibition downregulates expression of all core histones, implying a role in histone synthesis. Since histone synthesis occurs in parallel with DNA replication at S phase, we evaluated PCNA (Proliferating Cell Nuclear Antigen) expression, a protein involved in DNA replication and repair. PCNA expression was found to be significantly decreased in ANKHD1 silenced cells. We further observed accumulation γH2AX, a marker for DNA double stranded breaks and an early sign of DNA damage induced by replication stress, upon ANKHD1 silencing. The expressions of several genes implicated in DNA repair were also modulated in ANKHD1 silenced cells, confirming the role of ANKHD1 in DNA repair. Based on this study we speculate that ANKHD1 is an S phase protein required for histone synthesis and DNA repair. These results however, are preliminary and require thorough investigation.
Assuntos
Reparo do DNA , Histonas/metabolismo , Mieloma Múltiplo/metabolismo , Proteínas de Ligação a RNA/metabolismo , Linhagem Celular Tumoral , Replicação do DNA , Regulação Neoplásica da Expressão Gênica , Humanos , Mieloma Múltiplo/genética , Proteínas de Ligação a RNA/genética , Fase SRESUMO
Cancer still remains a major public health concern around the world and the search for new potential antitumor molecules is essential for fighting the disease. This study evaluated the anticancer and immunomodulatory potential of the newly synthetized ellipticine derivate: sodium bromo-5,11-dimethyl-6H-pyrido[4,3-b]carbazole-7-sulfonate (Br-Ell-SO3Na). It was prepared by the chlorosulfonation of 9-bromoellipticine. The ellipticine-7-sulfonic acid itself is not soluble, but its saponification with sodium hydroxide afforded a water-soluble sodium salt. The cytotoxicity of Br-Ell-SO3Na was tested against cancerous (K562 cell line) and non-cancerous cells (Vero cell line and human peripheral blood mononuclear cells (PBMC)) using a Methylthiazoletetrazolium (MTT) assay. Cell cycle arrest was assessed by flow cytometry and the immunomodulatory activity was analyzed through an enzyme-linked immunosorbent assay (ELISA). The results showed that the Br-Ell-SO3Na molecule has specific anticancer activity (IC50 = 35 µM) against the K562 cell line, once no cytotoxicity effect was verified against non-cancerous cells. Cell cycle analysis demonstrated that K562 cells treated with Br-Ell-SO3Na were arrested in the phase S. Moreover, the production of IL-6 increased and the expression of IL-8 was inhibited in the human PBMC treated with Br-Ell-SO3Na. The results demonstrated that Br-Ell-SO3Na is a promising anticancer molecule attested by its noteworthy activity against the K562 tumor cell line and immunomodulatory activity in human PBMC cells.
Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Elipticinas/química , Elipticinas/farmacologia , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Antineoplásicos/síntese química , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Técnicas de Química Sintética , Relação Dose-Resposta a Droga , Elipticinas/síntese química , Humanos , Fatores Imunológicos/síntese química , Imunomodulação/efeitos dos fármacos , Estrutura Molecular , Solubilidade , ÁguaRESUMO
BACKGROUND: The Duffy glycoprotein acts as the entry point for merozoites of Plasmodium vivax in the invasion of red blood cells. The host-parasite relationship has revealed new perspectives regarding the association between Duffy polymorphisms that can impact both the parasite density of this Plasmodium and the symptoms of this type of malaria. This study investigates the impact of Duffy polymorphisms on parasite density in patients infected with P. vivax in the Brazilian Amazon region. METHODS: Genotypes and Duffy polymorphism allele frequencies were compared in 287 patients with malaria, presenting low, medium and high density of P. vivax. The diagnosis of malaria was performed using a specialized team with a standardized clinical-laboratory method, while the Duffy genotyping was performed through the Bead Chip BioArray system. Both teams are reference services in Brazil. RESULTS: The FY*01 and FY*02 alleles were found in all three parasite density classes: low, medium and high, but when these alleles form genotypes with FY*02N.01 and FY*02W.01 alleles, they are found only in patients with low parasite density and low symptomatology. Another interesting finding found in this study is the presence of the genotype FY*02N.01/FY*02W.01 in one of the patients, presenting a very low parasite density and malaria considered subclinical, a genotype which had not been previously described in the literature. CONCLUSION: The presence of FY*02N.01 and FY*02W.01 alleles may have an impact on the reduction of clinical manifestations in malaria, leading to the development of subclinical malaria, making the infected individual an undetected natural reservoir, which may hinder the eradication of malaria in the Amazon.
Assuntos
Sistema do Grupo Sanguíneo Duffy/genética , Plasmodium vivax/fisiologia , Polimorfismo Genético , Adolescente , Adulto , Idoso , Brasil , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Densidade Demográfica , Adulto JovemRESUMO
OBJECTIVE/BACKGROUND: Sickle cell disease (SCD) is a chronic inflammatory condition caused by a point mutation in the HBB gene. Here we characterized the clinical presentation of SCD in a population from Amazonas State in northern Brazil, in order to evaluate whether the higher Amerindian ancestry observed in this relatively isolated geographic region would influence the clinical presentation of SCD. METHODS: This was a cross-sectional study characterizing the clinical presentation of SCD patients registered at HEMOAM, Amazon, Brazil. Data were obtained using a structured questionnaire, and by a review of the medical records. RESULTS: Of the 236 SCD patients listed in the historical records, 122 were included in this study. The median age was 15â¯years, with a male to female ratio of 52:70. The population was characterized by a high level of socioeconomic vulnerability, with only 2.1% presenting a family income above five minimum wages. Homozygous HbS (SS) was the most prevalent form of SCD (89.7%), and the diagnosis of SCD was performed in the context of complications in 92.3% of patients. The median frequency of vaso-occlusive crisis in the past 12â¯months was 2 (0-10). Using a validated clinical severity score based on clinical and laboratory data, no significant difference could be observed when compared to other populations. CONCLUSION: Our results represent the first comprehensive characterization of epidemiological, laboratorial, and clinical data of SCD in the region of the Brazilian Amazon. Despite the higher contribution of Amerindian ancestry previously demonstrated in this region, the main clinical characteristics of SCD seem similar to those reported in other populations.
Assuntos
Anemia Falciforme/epidemiologia , Inquéritos e Questionários , Adolescente , Adulto , Brasil/epidemiologia , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Índice de Gravidade de DoençaRESUMO
Chryseobacterium indologenes is an emerging nosocomial pathogen that produces IND-type chromosomal metallo-beta-lactamase. The phenotype and molecular aspects of two multidrug resistant C. indologenes strains and the analysis of the tertiary structure of the IND enzyme were studied. Identification of species and susceptibility tests were performed using the Vitek-2 compact. Chromosomal and plasmid DNA were extracted using PureLink™ Genomic DNA Mini Kit and PureLink Quick Plasmid Miniprep Kit, and the sequencing was performed using ABI 3130 genetic analyzer. Two strains were isolated and are registered as P-23 and P-113. Of the two, P-113 was sensitive to ciprofloxacin and cefepime only, whereas the P-23 showed reduced sensitivity to ceftazidime, ciprofloxacin, and tigecycline. The genetic analysis of both isolates identified the presence of the blaIND-like gene, with similarity to IND-3 and IND-8 alleles. The IND-3 identified in the P-133 sample presented a single mutation at position T355G, which corresponds to a nonsynonymous substitution of the amino acid at position 119 (SerâAla). The phylogenetic analysis of INDs showed lineages that are circulating in Asian and European countries. These results emphasize the need for effective preventive actions to avoid the dissemination of this type of pathogen in the hospital environment.
Assuntos
Antibacterianos/farmacologia , Chryseobacterium/genética , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Flavobacteriaceae/microbiologia , beta-Lactamases/genética , Idoso de 80 Anos ou mais , Substituição de Aminoácidos , Brasil , Cefepima/farmacologia , Ceftazidima/farmacologia , Cromossomos Bacterianos/química , Cromossomos Bacterianos/metabolismo , Chryseobacterium/classificação , Chryseobacterium/efeitos dos fármacos , Chryseobacterium/isolamento & purificação , Ciprofloxacina/farmacologia , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/patologia , Feminino , Infecções por Flavobacteriaceae/tratamento farmacológico , Infecções por Flavobacteriaceae/patologia , Expressão Gênica , Humanos , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Modelos Moleculares , Filogenia , Plasmídeos/química , Plasmídeos/metabolismo , Mutação Puntual , Estrutura Secundária de Proteína , Tigeciclina/farmacologiaRESUMO
ANKHD1 (Ankyrin repeat and KH domain-containing protein 1) is highly expressed and plays an important role in the proliferation and cell cycle progression of multiple myeloma (MM) cells. ANKHD1 downregulation modulates cell cycle gene expression and upregulates p21 irrespective of the TP53 mutational status of MM cell lines. The present study was aimed to investigate the role of ANKHD1 in MM in vitro clonogenicity and in vivo tumourigenicity, as well as the role of ANKHD1 in p21 transcriptional regulation. ANKHD1 silencing in MM cells resulted in significantly low no. of colonies formed and in slow migration as compared to control cells (p < 0.05). Furthermore, in xenograft MM mice models, tumour growth was visibly suppressed in mice injected with ANKHD1 silenced cells compared to the control group. There was a significant decrease in tumour volume (p = 0.006) as well as in weight (p = 0.02) in the group injected with silenced cells compared to those of the control group. Co-immunoprecipitation and chromatin immunoprecipitation (ChIP) assays confirmed the interaction between p21 and ANKHD1. Moreover, overexpression of ANKHD1 downregulated the activity of a p21 promoter in luciferase assays. Decrease in luciferase activity suggests a direct role of ANKHD1 in p21 transcriptional regulation. In addition confocal analysis after U266 cells were treated with Leptomycin B (LMB) for 24 h showed accumulation of ANKHD1 inside the nucleus as compared to untreated cells where ANKHD1 was found to be predominantly in cytoplasm. This suggests ANKHD1 might be shuttling between cytoplasm and nucleus. In conclusion, ANKHD1 promotes MM growth by repressing p21 a potent cell cycle regulator.
Assuntos
Proliferação de Células , Inibidor de Quinase Dependente de Ciclina p21/genética , Mieloma Múltiplo/genética , Regiões Promotoras Genéticas/genética , Proteínas de Ligação a RNA/genética , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Animais , Western Blotting , Linhagem Celular Tumoral , Movimento Celular/genética , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Ácidos Graxos Insaturados/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica , Células HEK293 , Humanos , Masculino , Camundongos Endogâmicos NOD , Camundongos SCID , Microscopia Confocal , Mieloma Múltiplo/metabolismo , Mieloma Múltiplo/patologia , Ligação Proteica , Interferência de RNA , Proteínas de Ligação a RNA/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Carga Tumoral/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
ANKHD1 is a multiple ankyrin repeat containing protein, highly expressed in cancers, such as acute leukemia. The present study was undertaken to determine the expression and functional significance of ANKHD1 in human Multiple Myeloma (MM). We found that ANKHD1 is highly expressed in MM patient cells and cell lines. In vitro, lentiviral mediated ANKHD1-shRNA inhibited proliferation and delayed S to G2M cell cycle progression in glucocorticoid resistant (U266) and sensitive (MM1S) MM cells. Further ANKHD1 silencing resulted in upregulation of cyclin dependent kinase inhibitor p21 irrespective of the p53 status of the MM cell lines. These data suggest that ANKHD1 might have a role in MM cell proliferation and cell cycle progression by regulating expression of p21.
Assuntos
Ciclo Celular , Proliferação de Células , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Mieloma Múltiplo/metabolismo , Proteínas de Ligação a RNA/biossíntese , Linhagem Celular Tumoral , Inativação Gênica , Glucocorticoides/uso terapêutico , Humanos , Mieloma Múltiplo/genética , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteínas de Ligação a RNA/genética , Proteína Supressora de Tumor p53/metabolismo , Regulação para CimaRESUMO
Pollen from the mesquite tree, Prosopis juliflora, is an important source of respiratory allergy in tropical countries. Our aim was to partially characterize the IgE binding proteins of P. juliflora pollen extract and study cross-reactivity with prevalent tree pollen allergens. Intradermal tests with P. juliflora and five other tree pollen extracts were performed on respiratory allergy patients from Bikaner (arid) and Delhi (semi arid). Prosopis extract elicited positive skin reactions in 71/220 of the patients. Sera were collected from 38 of these 71 patients and all demonstrated elevated specific IgE to P. juliflora. Immunoblotting with pooled patients' sera demonstrated 16 IgE binding components, with components of 24, 26, 29, 31, 35, 52, 58, 66 and 95 kDa recognized by more than 80% of individual patients' sera. P. juliflora extract is allergenically potent requiring 73 ng of self-protein for 50% inhibition of IgE binding in ELISA inhibition. Cross-inhibition assays showed close relationship among P. juliflora, Ailanthus excelsa, Cassia siamea and Salvadora persica. IgE binding components of 14, 41, 52 and 66 kDa were shared allergens whereas 26 and 29 kDa were specific to P. juliflora. The findings suggest that purification of cross-reactive allergens will be helpful for diagnosis and immunotherapy of tree pollen allergic patients.