RESUMEN
PURPOSE: The GEO database and KEGG database-based analyses identified the differential expression of cyclin-dependent kinase 1 (CDK1) in cervical cancer and its involvement in the cell cycle pathway. In the present study, we aim to clarify the role of CDK1 in cervical cancer and the function of upstream microRNA (miR)-143-3p/miR-495-3p. METHODS: The expression of miR-143-3p, miR-495-3p, and CDK1 in cervical cancer tissues and cells was determined using RT-qPCR. Cell bioactivities were examined by CCK-8 and flow cytometry. The binding affinity between CDK1 and miR-143-3p/miR-495-3p was investigated using dual luciferase gene reporter assay. A xenograft mouse model of cervical cancer was then established to explore their effect on the tumorigenicity of cervical cancer cells in vivo. RESULTS: CDK1 was found to be the common target gene of miR-143-3p and miR-495-3p. CDK1 overexpression occurred in cervical cancer tissues and cells, while expression of miR-495-3p and miR-143-3p was down-regulated. The viability was inhibited while the apoptosis was promoted in cervical cancer cells in response to miR-143-3p or miR-495-3p overexpression, or CDK1 silencing. Further, miR-143-3p or miR-495-3p overexpression was also substantiated to inhibit the tumorigenicity of cervical cancer cells in vivo, while CDK1 overexpression counteracted their effect. CONCLUSION: Taken together, miR-143-3p and miR-495-3p co-target CDK1, thereby inhibiting the occurrence and development of cervical cancer.
Asunto(s)
Proteína Quinasa CDC2/metabolismo , MicroARNs/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Animales , Apoptosis , Proteína Quinasa CDC2/genética , Ciclo Celular , Movimiento Celular , Proliferación Celular , Cuello del Útero/metabolismo , Bases de Datos Genéticas , Regulación hacia Abajo , Femenino , Silenciador del Gen , Genes Reporteros , Células HeLa , Xenoinjertos , Humanos , Luciferasas/genética , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Invasividad Neoplásica , Trasplante de Neoplasias , Distribución Aleatoria , Regulación hacia Arriba , Neoplasias del Cuello Uterino/patologíaRESUMEN
The Coatomer protein complex subunit beta 2 (COPB2) is involved in the formation of the COPI coatomer protein complex and is responsible for the transport of vesicles between the Golgi apparatus and the endoplasmic reticulum. It plays an important role in maintaining the integrity of these cellular organelles, as well as in maintaining cell homeostasis. More importantly, COPB2 plays key roles in embryonic development and tumor progression. COPB2 is regarded as a vital oncogene in several cancer types and has been implicated in tumor cell proliferation, survival, invasion, and metastasis. Here, we summarize the current knowledge on the roles of COPB2 in cancer development and progression in the context of the hallmarks of cancer.
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Proteína Coatómero/fisiología , Neoplasias/etiología , Animales , Apoptosis/genética , Apoptosis/fisiología , Muerte Celular Autofágica/fisiología , Ciclo Celular/fisiología , Proliferación Celular/genética , Supervivencia Celular/genética , Proteína Coatómero/genética , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Desarrollo Embrionario , Retículo Endoplásmico/fisiología , Aparato de Golgi/fisiología , Homeostasis , Humanos , Ratones , Invasividad Neoplásica/genética , Invasividad Neoplásica/fisiopatología , Metástasis de la Neoplasia/genética , Metástasis de la Neoplasia/fisiopatología , Neoplasias/patología , Vesículas Transportadoras/fisiologíaRESUMEN
Although therapeutic hypothermia is an effective treatment for post-resuscitation brain injury after cardiac arrest (CA), the underlying mechanism remains unclear. Vacuolar H(+)-ATPase (V-ATPase) plays a key role in cellular adaption to a hypoxic environment. This study sought to evaluate the effect of mild hypothermia on V-ATPase and its involvement in neuroprotection after CA. Male Sprague-Dawley rats were subjected to a 6-min CA, resuscitated successfully, and then assigned to either the normothermia (NT) group or the hypothermia (HT) group. Rats were further divided into 2 subgroups based on the time of euthanasia, either 3 or 24 h after CA (NT-3 h, HT-3 h; NT-24 h, HT-24 h). Mild hypothermia was induced following CA and maintained at 33°C for 2 h. Neurologic deficit scores were used to determine the status of neurological function. Brain specimens were analyzed by TUNEL assay, western blotting, and immunohistochemistry. V-ATPase activity was estimated by subtracting total ATP hydrolysis from the bafilomycin-sensitive activity. Mild hypothermia improved the neurological outcome (HT-24 h: 34.3 ± 16.4 vs NT-24 h: 50.3 ± 17.4) and significantly decreased neurocyte apoptosis 24 h after resuscitation. Mild hypothermia significantly increased V0a1 compared to NT-3 h; V0a1 expression was associated with a decrease in the cleaved caspase 3 expression. These findings suggested that mild hypothermia inhibits CA-induced apoptosis in the hippocampus, which may be associated with reduced V-ATPase impairment. These data provide new insights into the protective effects of hypothermia in vivo.
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Lesiones Encefálicas/terapia , Paro Cardíaco/terapia , Hipotermia Inducida , ATPasas de Translocación de Protón Vacuolares/biosíntesis , Animales , Apoptosis/genética , Encéfalo/metabolismo , Encéfalo/patología , Lesiones Encefálicas/etiología , Lesiones Encefálicas/genética , Lesiones Encefálicas/patología , Caspasa 3/biosíntesis , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/genética , Paro Cardíaco/complicaciones , Paro Cardíaco/metabolismo , Paro Cardíaco/patología , Humanos , Masculino , Ratas , Resucitación , ATPasas de Translocación de Protón Vacuolares/genética , ATPasas de Translocación de Protón Vacuolares/aislamiento & purificaciónRESUMEN
Fruit ripening is a complex developmental process, the details of which remain largely unknown in fleshy fruits. In this paper, the fruit flesh of two peach varieties, "Zhongyou9" (a nectarine; Prunus persica L. Batsch) and its mutant "Hongyu", was analyzed by RNA-seq technology during two stages of ripening at 20-day intervals. One hundred and eighty significant upregulated and two hundred and thirty-five downregulated genes were identified in the experiment. Many of these genes were related to plant hormones, chlorophyll breakdown, accumulation of aroma and flavor volatiles, and stress. To the best of our knowledge, this is the first transcriptome analysis of peach ripening, and our data will be useful for further studies of the molecular basis of fruit ripening.
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Frutas/genética , Perfilación de la Expresión Génica , Prunus persica/genética , Transcriptoma , Regulación de la Expresión Génica de las Plantas , Mutación , Prunus persica/metabolismoRESUMEN
With the development of chrysanthemum breeding in recent years, an increasing number of wild species in genera related to Chrysanthemum were introduced to extend the genetic resources and facilitate the genetic improvement of chrysanthemums via hybridization. However, few simple sequence repeat (SSR) markers are available for marker-assisted breeding and population genetic studies of chrysanthemum and closely related species. Expressed sequence tags (ESTs) in public databases and cross-species transferable markers are considered to be a cost-effective means for developing sequence-based markers. In this study, 25 EST-SSRs were successfully developed from Chrysanthemum EST sequences for Chrysanthemum morifolium and closely related species. In total, 4164 unigene sequences were assembled from 7180 ESTs of chrysanthemum in GenBank, which were subsequently used to screen for the presence of microsatellites with the SSRIT software. The screening criteria were 8, 5, 4, and 3 repeating units for di-, tri-, tetra-, and penta- and higher-order nucleotides, respectively. Moreover, 310 SSR loci from 296 sequences were identified, and 198 primer pairs for SSR amplification were designed with the Primer Premier 5.0 software, of which 25 SSR loci showed polymorphic amplification in 52 species and varieties belonging to Chrysanthemum, Ajania, and Opisthopappus. The application of EST-SSR markers to the identification of intergeneric hybrids between Chrysanthemum and Ajania was demonstrated. Therefore, EST-SSRs can be developed for species that lack gene sequences or ESTs by utilizing ESTs of closely related species.
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Chrysanthemum/genética , Etiquetas de Secuencia Expresada/metabolismo , Repeticiones de Microsatélite/genética , Filogenia , Marcadores Genéticos , Hibridación GenéticaRESUMEN
Plant height is one of the most important traits of plant architecture as it modulates both economic and ornamental values. Crape myrtle (Lagerstroemia indica L.) is a popular ornamental woody plant because of its long-lasting mid-summer bloom, rich colors, and diversified plant architecture. These traits also make it an ideal model of woody species for genetic analysis of many ornamental traits. To understand the inheritance of plant height and screen for genes modulating plant height in Lagerstroemia, segregation of the plant height trait was analyzed using the F1 population of L. fauriei (standard) x L. indica 'Pocomoke' (dwarf) with 96 seedlings, while dwarf genes were screened using the bulked segregant analysis method, combined with 28 amplified fragment length polymorphism primers and 41 simple sequence repeat primers. The results showed that the dwarf trait of crape myrtle was controlled by a major gene and modified by minor genes. An amplified fragment length polymorphism marker, M53E39-92, which was 23.33 cM from the loci controlling the dwarf trait, was screened. These results provide basic information for marker-assisted selection in Lagerstromia and cloning of dwarf genes in future studies.
Asunto(s)
Genes de Plantas , Lagerstroemia/anatomía & histología , Fenotipo , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Biometría , Marcadores Genéticos , Lagerstroemia/genéticaRESUMEN
The entomopathogenic fungus Verticillium lecanii is a well-known biocontrol agent of fungal phytopathogens, as well as insect pests. A 42-kDa chitinase belonging to family 18 of the glycosyl hydrolases was isolated and partially characterized. Chitinase was purified using successive column chromatography on phenyl-sepharose, DEAE-sepharose, and CM-sepharose. The enzyme showed the highest activity at 40°C and pH 4.6. Enzyme activity was strongly activated in the presence of Mg(2+). The purified enzyme showed inhibitory activity of spore germination against several plant pathogens, particularly Fusarium moniliforme. The genomic DNA and cDNA sequences were resolved by polymerase chain reaction amplification and sequencing. Protein modeling and comparative investigation of different chitinase amino acids showed that chitinases are conserved in parasitic fungi.
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Quitinasas/genética , Proteínas Fúngicas/genética , Insectos/microbiología , Verticillium/genética , Animales , Antifúngicos/farmacología , Quitinasas/aislamiento & purificación , Quitinasas/metabolismo , Cromatografía/instrumentación , Cromatografía/métodos , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Estabilidad de Enzimas , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/farmacología , Fusarium/efectos de los fármacos , Fusarium/crecimiento & desarrollo , Concentración de Iones de Hidrógeno , Cinética , Datos de Secuencia Molecular , Sefarosa , Análisis de Secuencia de ADN , Esporas Fúngicas/efectos de los fármacos , Esporas Fúngicas/crecimiento & desarrollo , Temperatura , Verticillium/enzimologíaRESUMEN
Simple, efficient, and economical recombinant plant binary expression vectors for deciphering large-scale functional genomic research in plants and promoting crop improvement by genetically engineering and biotechnology is in great demand. In this research, using the pCHF3, pCAMBIA1301, pCAMBIA3300, pCAMBIA3301 vectors, we successfully constructed general plant binary expression vectors carrying CaMV35S and Arabidopsis rd29A promoters mediating multiple cloning sites ( MCS: SacI, KpnI, SmaI, BamHI, XbaI, SalI, and PstI). Meanwhile, a series of applicative binary expression vectors that can be utilized for subcellular localization were constructed by fusion of the MCS and eGFP. Subsequently, the recombinant vectors were successfully transferred into Arabidopsis thaliana and Nicotiana benthamiana for further investigation of functional elements in these plant binary expression vectors. Our results demonstrated that this system was a convenient and versatile vector system for phenotypic, functional, subcellular localization, and promoter activity analysis, and it provided a relatively high-efficiency and reliable platform for researchers in vector construction and may facilitate large-scale functional genomics analysis in plants.
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Vectores Genéticos , Genoma de Planta , Secuencia de Bases , Cartilla de ADN , Genes Reporteros , Glucuronidasa/genética , Proteínas Fluorescentes Verdes/genética , Reacción en Cadena de la Polimerasa , Fracciones Subcelulares/metabolismoRESUMEN
The cDNA sequence of foot-specific peroxidase PPOD1 from the Chinese strain of Hydra magnipapillata was cloned by reverse transcription-polymerase chain reaction. The cDNA sequence contained a coding region with an 873-bp open reading frame, a 31-bp 5'-untranslated region, and a 36-bp 3'-untranslated region. The structure prediction results showed that PPOD1 contains 10.34% of α-helix, 38.62% of extended strand, 12.41% of ß-turn, and 38.62% of random coil. The structural core was α-helix at the N terminus. The GenBank protein blast server showed that PPOD1 contains 2 fascin-like domains. In addition, high-level PPOD1 activity was only present in the ectodermal epithelial cells located on the edge of the adhesive face of the basal disc, and that these cells extended lamellipodia and filopodia when the basal disc was tightly attached to a glass slide. The fascin-like domains of Hydra PPOD1 might contribute to the bundling of the actin filament of these cells, and hence, the formation of filopodia. In conclusion, these cells might play an important role in strengthening the adsorbability of the basal disc to substrates.
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Regulación Enzimológica de la Expresión Génica , Hydra/genética , Sistemas de Lectura Abierta/genética , Peroxidasa/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Sitios de Unión/genética , China , Clonación Molecular , ADN Complementario/química , ADN Complementario/genética , Hydra/enzimología , Modelos Moleculares , Datos de Secuencia Molecular , Peroxidasa/clasificación , Peroxidasa/metabolismo , Filogenia , Células Procariotas/metabolismo , Estructura Terciaria de Proteína , Seudópodos/enzimología , Seudópodos/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de Secuencia de AminoácidoRESUMEN
Evidence suggests that some genetic variants are risk factors for both colorectal cancer (CRC) and gastric cancer (GC). Thus, we selected 12 reported single nucleotide polymorphisms (SNPs) from genome-wide association studies of CRC and conducted this case-control study to assess the associations between these SNPs and the risk for GC in a southern Chinese population. All SNPs were genotyped in 249 individuals with GC and 292 healthy population-matched subjects using the Sequenom MassArray iPLEX System. Association analyses based on the c2 test and binary logistic regression were performed to determine the odds ratio (OR) and 95% confidence interval (95%CI) for each SNP. A stratified analysis by gender was also performed. Borderline significant associations were observed for rs4444235 (P = 0.070) and rs10411210 (P = 0.084), both fitting the overdominant model. The rs4444235 CT genotype showed a protective effect (OR = 0.72, 95%CI = 0.50-1.03), while the rs10411210 CT genotype was a risk factor (OR = 1.40, 95%CI = 0.96-2.05) as compared with the CC+TT genotype. In the female subgroup, the rs6983267 GT genotype (compared with TT, OR = 2.31, 95%CI = 1.07-4.99) and the rs10505477 CT genotype (compared with TT, OR = 2.36, 95%CI = 1.09-5.11) significantly increased the risk for GC. No significant association was detected for the other SNPs. These results provide evidence that known genetic variants associated with CRC risk may also confer risk for GC.
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Neoplasias Colorrectales/genética , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Neoplasias Gástricas/genética , Adulto , Anciano , Estudios de Casos y Controles , Neoplasias Colorrectales/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Factores de Riesgo , Caracteres Sexuales , Neoplasias Gástricas/patologíaRESUMEN
Lagerstroemia (crape myrtle) are famous ornamental plants with large pyramidal racemes, long flower duration, and diverse colors. However, little is known about the genetic structure and diversity of germplasm in Lagerstroemia. We genotyped 81 L. indica cultivars, five other species of Lagerstroemia, and 10 interspecific hybrids using 30 simple sequence repeat markers; 275 alleles were generated with a mean of nine alleles per locus. The mean polymorphism information content value, a measure of gene diversity, was 0.63, with a range from 0.25 to 0.86. The mean observed heterozygosity (0.51) tended to be lower than the mean expected heterozygosity (0.67). The mean F-statistics (F(ST), F(IS), and F(IT)) were 0.05, 0.20, and 0.24, respectively, indicating a high level of genetic variation among cultivars. Clustering analysis based on genetic distance divided the 96 genotypes into three distinct groups, which corresponded with their genetic backgrounds and geographic regions. L. indica cultivars and the other five L. species were grouped into different sub-clusters. Chinese and North American cultivars were divided into different clusters. These data about the genetic relationship among cultivars demonstrated the potential value of L. indica cultivars and other Lagerstroemia species for widening the genetic basis of breeding programs for this ornamental flower.
Asunto(s)
Variación Genética , Lagerstroemia/genética , Repeticiones de Microsatélite/genética , Alelos , Análisis por Conglomerados , Sitios Genéticos/genética , Marcadores Genéticos , Especificidad de la EspecieRESUMEN
Patterns of DNA methylation are established and maintained by a family of DNA methyltransferases (DNMTs). Aberrant promoter DNA methylation of tumor suppressor genes is found in breast cancer. Association studies between DNMT gene polymorphisms and breast cancer in various populations have reported inconsistent results. This study assessed the associations of single nucleotide polymorphisms (SNPs) in DNMT1, DNMT3A, DNMT3B, DNMT3L, and DNMT2 with breast cancer among Han Chinese women from South China. Sixteen SNPs (rs2114724, rs2228611, rs2228612, rs8101866, and rs16999593 in DNMT1; rs13420827, rs11887120, rs13428812, rs1550117, rs11695471, and rs6733301 in DNMT3A; rs2424908, rs2424913, and rs6087990 in DNMT3B; rs113593938 in DNMT3L, and rs11254413 in DNMT2) in 408 women with breast cancer and 469 controls were genotyped using a MassARRAY matrix-assisted laser desorption/ionization time-of-flight mass spectrometry platform. Two SNPs, rs16999593 in DNMT1 and rs2424908 in DNMT3B, were significantly associated with breast cancer risk. The heterozygous genotype CT of rs16999593 was associated with increased breast cancer risk [odds ratio (OR) = 1.60; 95% confidence interval (95%CI) = 1.20-2.14; P = 0.0052], whereas rs2424908 was associated with decreased risk (OR = 0.62; 95%CI = 0.46-0.84; P = 0.0061). Other DNMT polymorphisms showed no significant associations with breast cancer risk in the study population. Haplotype CGTC of rs2114724, rs2228611, rs8101866, and rs16999593 in DNMT1 differed significantly as a risk factor between the case and control groups (OR = 1.51; 95%CI = 1.18-1.93; P = 0.0012). The heterozygous genotypes of rs16999593 in DNMT1 and rs2424908 in DNMT3B were strongly associated with breast cancer risk.
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Neoplasias de la Mama/genética , ADN (Citosina-5-)-Metiltransferasas/genética , Polimorfismo de Nucleótido Simple , Estudios de Casos y Controles , China , ADN (Citosina-5-)-Metiltransferasa 1 , Femenino , Frecuencia de los Genes , Genes Dominantes , Estudios de Asociación Genética , Genotipo , Humanos , Desequilibrio de Ligamiento , Persona de Mediana Edad , Oportunidad Relativa , Factores de Riesgo , Análisis de Secuencia de ADN , ADN Metiltransferasa 3BRESUMEN
A segurança da pravastatina, um inibidor seletivo hidrofílico da HMG-CoA redutase, foi avaliada em 1.142 pacientes adultos hipercolesterolêmicos em seis ensaios clínicos multicêntricos realizados nos Estados Unidos. Todos os ensaios foram aleatórios, duplo-cegos e controlados por placebo ao menos nas primeiras oito a 16 semanas, mantendo-se a seguir o tratamento a longo prazo com pravastatina em esquema duplo-cego ou aberto, com ou sem outras drogas redutoras de lipídios; o controle ativo primário foi a resina biliar de ligaçao a ácidos. Nao houve diferenças entre os grupos de pravastatina e placebo com relaçao às taxas globais de reaçoes adversas atribuíveis à droga. Somente erupçoes cutâneas (rash) ocorreram com maior freqúência estatística (p < 0,01) nos pacientes tratados com pravastina. Essas erupçoes geralmente eram leves e transitórias e somente 1,3 por cento dos casos relatados em pacientes tratados com pravastatina estavam possivelmente relacionados à terapia. Durante um período médio de tratamento de 18 meses, as razoes mais freqüentes para a interrupçao da administraçao de pravastatina foram queixas abdominais leves (1,4 por cento dos pacientes) e aumentos assintomáticos das transaminases hepáticas (1,0 por cento dos pacientes). Os valores médios de ALAT e ASAT aumentaram ligeiramente, atingindo um patamar estável após as primeiras oito semanas de terapia. Nenhuma ocorrência de enzimas hepáticas anormais entre os pacientes tratados com pravastina foi associada a doença clínica. Aumentos semelhantes das transaminases hepáticas também foram observados nos pacientes tratados com resina. Em geral, a pravastatina foi bem tolerada e aparentemente nao afetou os músculos esqueléticos, o sono ou o desenvolvimento de catarata.