RESUMO
Berberine (BBR) is a natural alkaloid with significant anti-tumor activity against many types of cancer cells. In this study, we investigated the molecular mechanisms employed by BBR to repress the proliferation and growth of skin squamous cell carcinoma A431 cells. Berberine was reported to inhibit the proliferation of A431 cells in a dose- and time-dependent manner and was observed to induce a series of biochemical events, including the loss of mitochondrial membrane potential, release of cytochrome-c to cytosol, induction of proteins of the Bcl-2 family and caspases, and the cleavage of poly(ADP)-ribose polymerase. This suggested its ability to induce apoptosis. The results of a wound healing test revealed that berberine inhibited the migration of A431 cells. Ezrin was transfected into A431 cells by RNA interference. The level of expression of Ezrin in the transfected A431 cells was observed to decrease with berberine treatment, which suggested that berberine might inhibit the invasion of A431 cells through Ezrin. The results of this study demonstrated that berberine could potentially inhibit proliferation, induce apoptosis, and inhibit the invasion of A431 cells.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Berberina/farmacologia , Proteínas do Citoesqueleto/genética , Células Epiteliais/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citocromos c/genética , Citocromos c/metabolismo , Proteínas do Citoesqueleto/metabolismo , Relação Dose-Resposta a Droga , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Poli(ADP-Ribose) Polimerases/genética , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais , Pele/efeitos dos fármacos , Pele/metabolismo , Pele/patologia , Transgenes , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
The purpose of this study was to identify differentially expressed genes and analyze biological processes related to leukemia. A meta-analysis was performed using the Rank Product package of Gene Expression Omnibus datasets for leukemia. Next, Gene Ontology-enrichment analysis and pathway analysis were performed using the Gene Ontology website and Kyoto Encyclopedia of Genes and Genomes. A protein-protein interaction network was constructed using the Cytoscape software. Using the Rank Product package for leukemia, we identified a total of 1294 differentially expressed genes, 357 of which were not involved in individual differentially expressed genes. Gene Ontology-enrichment analyses showed that these 357 genes were enriched in biological processes such as mRNA metabolism, RNA splicing, and mRNA processing. Pathway-enrichment analysis showed that the genes were involved in the intestinal immune network for IgA production, endocytosis, and the mitogen-activated protein kinase signaling pathway. The protein-protein interaction network indicated that HRAS, CD44, STAT1, SMAD2, and COPS5 were important in many interactions. Our study revealed genes that were consistently differentially expressed in leukemia, as well as the biological pathways and protein-protein interaction network associated with these genes.
Assuntos
Regulação Leucêmica da Expressão Gênica/genética , Leucemia/genética , Proteínas de Neoplasias/biossíntese , Análise de Sequência com Séries de Oligonucleotídeos , Bases de Dados Genéticas , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Leucemia/patologia , Proteínas Quinases Ativadas por Mitógeno/biossíntese , Transdução de Sinais/genética , SoftwareRESUMO
Diabetics have an increased prevalence of periodontitis, and diabetes is one of the causative factors of severe periodontitis. Apoptosis is thought to be involved in this pathogenic relationship. The aim of this study was to investigate apoptosis in human periodontal ligament (PDL) fibroblasts induced by advanced glycation end products (AGEs) and their receptor (RAGE). We examined the roles of apoptosis, AGEs, and RAGE during periodontitis in diabetes mellitus using cultured PDL fibroblasts that were treated by AGE-modified bovine serum albumin (AGE-BSA), bovine serum albumin (BSA) alone, or given no treatment (control). Microscopy and real-time quantitative PCR indicated that PDL fibroblasts treated with AGE-BSA were deformed and expressed higher levels of RAGE and caspase 3. Cell viability assays and flow cytometry indicated that AGE-BSA reduced cell viability (69.80±5.50%, P<0.01) and increased apoptosis (11.31±1.73%, P<0.05). Hoechst 33258 staining and terminal-deoxynucleotidyl transferase-mediated nick-end labeling revealed that AGE-BSA significantly increased apoptosis of PDL fibroblasts. The results showed that the changes in PDL fibroblasts induced by AGE-BSA may explain how AGE-RAGE participates in and exacerbates periodontium destruction.
Assuntos
Animais , Bovinos , Humanos , Apoptose/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , /farmacologia , Ligamento Periodontal/citologia , Receptores Imunológicos/metabolismo , Soroalbumina Bovina/farmacologia , Contagem de Células , /metabolismo , Sobrevivência Celular/efeitos dos fármacos , Complicações do Diabetes , Citometria de Fluxo , Fibroblastos/metabolismo , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Cultura Primária de Células , Doenças Periodontais/complicações , Ligamento Periodontal/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Diabetics have an increased prevalence of periodontitis, and diabetes is one of the causative factors of severe periodontitis. Apoptosis is thought to be involved in this pathogenic relationship. The aim of this study was to investigate apoptosis in human periodontal ligament (PDL) fibroblasts induced by advanced glycation end products (AGEs) and their receptor (RAGE). We examined the roles of apoptosis, AGEs, and RAGE during periodontitis in diabetes mellitus using cultured PDL fibroblasts that were treated by AGE-modified bovine serum albumin (AGE-BSA), bovine serum albumin (BSA) alone, or given no treatment (control). Microscopy and real-time quantitative PCR indicated that PDL fibroblasts treated with AGE-BSA were deformed and expressed higher levels of RAGE and caspase 3. Cell viability assays and flow cytometry indicated that AGE-BSA reduced cell viability (69.80 ± 5.50%, P<0.01) and increased apoptosis (11.31 ± 1.73%, P<0.05). Hoechst 33258 staining and terminal-deoxynucleotidyl transferase-mediated nick-end labeling revealed that AGE-BSA significantly increased apoptosis of PDL fibroblasts. The results showed that the changes in PDL fibroblasts induced by AGE-BSA may explain how AGE-RAGE participates in and exacerbates periodontium destruction.
Assuntos
Apoptose/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Produtos Finais de Glicação Avançada/farmacologia , Ligamento Periodontal/citologia , Receptores Imunológicos/metabolismo , Soroalbumina Bovina/farmacologia , Animais , Caspase 3/metabolismo , Bovinos , Contagem de Células , Sobrevivência Celular/efeitos dos fármacos , Complicações do Diabetes , Fibroblastos/metabolismo , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Doenças Periodontais/complicações , Ligamento Periodontal/efeitos dos fármacos , Cultura Primária de Células , Reação em Cadeia da Polimerase em Tempo Real , Receptor para Produtos Finais de Glicação AvançadaRESUMO
Genetic diversity of Elymus sibiricus (Poaceae) was examined in eight populations from the southeast Qinghai-Tibet Plateau. We detected 291 RAPD polymorphic loci in 93 samples. The percentage of polymorphic bands (PPB) was 79%. Genetic diversity (H(E)) was 0.264, effective number of alleles (N(E)) was 1.444, Shannon's information index (H(O)) was 0.398, and expected Bayesian heterozygosity (H(B)) was 0.371. At the population level, PPB = 51%, N(E) = 1.306, H(E) = 0.176, I = 0.263, and H(B) = 0.247. A high level of genetic differentiation was detected based on Nei's genetic diversity analysis (G(ST) = 32.0%), Shannon's index analysis (33.7%), and the Bayesian method (θ(B) = 33.5%). The partitioning of molecular variance by AMOVA demonstrated significant genetic differentiation within populations (60%) and among populations (40%). The average number of individuals exchanged between populations per generation (N(m)) was 1.06. The populations were found to share high levels of genetic identity. No significant correlation was found between geographic distance and pairwise genetic distance (r = 0.7539, P = 0.9996). Correlation analysis revealed a significant correlation (r = 0.762) between RAPD H(E) found in this study and ISSR H(E) values from a previous study.