RESUMO
PURPOSE: The aim of this study is to explore the roles of ß-catenin, decorin, septin-7, and S100A10 expression in colorectal cancer development. METHODS: Twenty-five BALB/c mice were divided into five groups; four groups were administrated N,N-dimethylhydrazine for 0, 10, 15, and 20 weeks, and one group was administrated normal saline for 20 weeks. The colons were collected for histopathological analysis. Protein samples prepared from the frozen colon tissues of mice treated with N,N-dimethylhydrazine for the different time points were evaluated using the isobaric tags for relative and absolute quantification (iTRAQ) labeling technique coupled with the 2D liquid chromatography-tandem mass spectrometry analysis. Based on the proteomic analysis results, immunohistochemical staining of ß-catenin, decorin, septin-7, and S100A10 was performed in paraffin-embedded mice colorectal tissue, and 53 cases of human hereditary polyposis colorectal cancer samples. RESULTS: Colorectal cancer was observed in mice treated with N,N-dimethylhydrazine for 20 weeks, and adenomas were observed in mice subjected to the 10-, and 15-week treatments. Seventy-two differentially expressed proteins were involved in the development of cancer as per the iTRAQ and spectrometry analysis. In normal epithelium, adenoma, and cancer from human hereditary polyposis colorectal cancer, S100A10 expression (c2 = 100.989, P = 0.000) was highest in cancer, whereas decorin (c2 = 12.852, P = 0.002) and septin-7 (c2 = 66.519, P = 0.002) expressions were highest in the normal epithelium, which was confirmed via immunohistochemical staining. CONCLUSIONS: The subcellular localization of ß-catenin and decorin, septin-7, and S100A10 expressions are associated with the development of colorectal cancer in mice after N,N-dimethylhydrazine treatment and in human hereditary polyposis colorectal cancers.
Assuntos
Polipose Adenomatosa do Colo/patologia , Biomarcadores Tumorais/análise , Neoplasias Colorretais/patologia , Adulto , Animais , Anexina A2/análise , Anexina A2/biossíntese , Carcinógenos/toxicidade , Neoplasias Colorretais/induzido quimicamente , Decorina/análise , Decorina/biossíntese , Dimetilidrazinas/toxicidade , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Proteômica/métodos , Proteínas S100/análise , Proteínas S100/biossíntese , Septinas/análise , Septinas/biossíntese , beta Catenina/análise , beta Catenina/biossínteseRESUMO
Cetuximab is widely used in patients with metastatic colon cancer expressing wildtype KRAS. However, acquired drug resistance limits its clinical efficacy. Exosomes are nanosized vesicles secreted by various cell types. Tumor cell-derived exosomes participate in many biological processes, including tumor invasion, metastasis, and drug resistance. In this study, exosomes derived from cetuximab-resistant RKO colon cancer cells induced cetuximab resistance in cetuximab-sensitive Caco-2 cells. Meanwhile, exosomes from RKO and Caco-2 cells showed different levels of phosphatase and tensin homolog (PTEN) and phosphor-Akt. Furthermore, reduced PTEN and increased phosphorylated Akt levels were found in Caco-2 cells after exposure to RKO cell-derived exosomes. Moreover, an Akt inhibitor prevented RKO cell-derived exosome-induced drug resistance in Caco-2 cells. These findings provide novel evidence that exosomes derived from cetuximab-resistant cells could induce cetuximab resistance in cetuximab-sensitive cells, by downregulating PTEN and increasing phosphorylated Akt levels.
Assuntos
Humanos , Neoplasias do Colo/tratamento farmacológico , PTEN Fosfo-Hidrolase/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Exossomos/efeitos dos fármacos , Cetuximab/farmacologia , Antineoplásicos Imunológicos/farmacologia , Sais de Tetrazólio , Fatores de Tempo , Western Blotting , Análise de Variância , Células CACO-2 , Linhagem Celular TumoralRESUMO
Cetuximab is widely used in patients with metastatic colon cancer expressing wildtype KRAS. However, acquired drug resistance limits its clinical efficacy. Exosomes are nanosized vesicles secreted by various cell types. Tumor cell-derived exosomes participate in many biological processes, including tumor invasion, metastasis, and drug resistance. In this study, exosomes derived from cetuximab-resistant RKO colon cancer cells induced cetuximab resistance in cetuximab-sensitive Caco-2 cells. Meanwhile, exosomes from RKO and Caco-2 cells showed different levels of phosphatase and tensin homolog (PTEN) and phosphor-Akt. Furthermore, reduced PTEN and increased phosphorylated Akt levels were found in Caco-2 cells after exposure to RKO cell-derived exosomes. Moreover, an Akt inhibitor prevented RKO cell-derived exosome-induced drug resistance in Caco-2 cells. These findings provide novel evidence that exosomes derived from cetuximab-resistant cells could induce cetuximab resistance in cetuximab-sensitive cells, by downregulating PTEN and increasing phosphorylated Akt levels.
Assuntos
Antineoplásicos Imunológicos/farmacologia , Cetuximab/farmacologia , Neoplasias do Colo/tratamento farmacológico , Exossomos/efeitos dos fármacos , PTEN Fosfo-Hidrolase/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Análise de Variância , Western Blotting , Células CACO-2 , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/metabolismo , Resistência a Medicamentos , Exossomos/metabolismo , Formazans , Humanos , PTEN Fosfo-Hidrolase/análise , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/análise , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sais de Tetrazólio , Fatores de TempoRESUMO
The objective of this study was to use RNA interference (RNAi) to improve protein quality and decrease anti-nutritional effects in soybean. Agrobacterium tumefaciens-mediated transformation was conducted using RNAi and an expression vector containing the 7S globulin ß-subunit gene. The BAR gene was used as the selective marker and cotyledonary nodes of soybean genotype Jinong 27 were chosen as explant material. Regenerated plants were detected by molecular biology techniques. Transformation of the ß-subunit gene in the 7S protein was detected by PCR, Southern blot, and q-PCR. Positive plants (10 T0, and 6 T1, and 13 T2) were tested by PCR. Hybridization bands were detected by Southern blot analysis in two of the T1 transgenic plants. RNAi expression vectors containing the soybean 7S protein ß-subunit gene were successfully integrated into the genome of transgenic plants. qRT-PCR analysis in soybean seeds showed a clear decrease in expression of the soybean ß-subunit gene. The level of 7S protein ß-subunit expression in transgenic plants decreased by 77.5% as compared to that of the wild-type plants. This study has established a basis for the application of RNAi to improve the anti-nutritional effects of soybean.
Assuntos
Agrobacterium tumefaciens/genética , Antígenos de Plantas/genética , Globulinas/genética , Glycine max/genética , Interferência de RNA , Proteínas de Armazenamento de Sementes/genética , Proteínas de Soja/genética , Antígenos de Plantas/metabolismo , Cotilédone/citologia , Cotilédone/genética , Cotilédone/metabolismo , Técnicas de Transferência de Genes , Genoma de Planta , Globulinas/metabolismo , Recombinação Genética , Proteínas de Armazenamento de Sementes/metabolismo , Proteínas de Soja/metabolismo , TransgenesRESUMO
The susceptibility to glioma is not well understood. It has been suggested that the X-ray cross complementing group 3 (XRCC3) gene influences the capacity to repair DNA damage, leading to increased glioma susceptibility. In this study, we evaluated the relationship between XRCC3 mutations and glioma risk. Genotypes were assessed in 389 Chinese glioma patients and 358 healthy controls. XRCC3 Thr241Met (rs861539) and 2 additional polymorphisms, rs3212112 (c.774+19T>G) and rs1799796 (c.562-14A>G), were directly sequenced. The frequency of the rs861539 T allele was significantly lower in the glioma group than in healthy controls [11.1 vs 17.7%, odds ratio = 0.62 (0.48-0.80), P < 0.001]; the frequencies of the CT or CT+TT genotypes differed between groups (18.5 vs 31%, 20.3 vs 33.2%, respectively). The frequency of the rs3212112 G allele was significantly higher in the glioma group than in healthy controls [15.8 vs 5.3%, odds ratio = 2.94 (2.07-4.17), P < 0.001]. The frequencies of the GT or TG+GG genotypes differed between groups (25.4 vs 7.8%, 28.5 vs 9.2%, respectively). This study demonstrates that the rs861539 and rs3212112 polymorphisms in the XRCC3 gene may influence the risk of glioma development in Chinese populations.
Assuntos
Neoplasias Encefálicas/genética , Reparo do DNA , Proteínas de Ligação a DNA/genética , Predisposição Genética para Doença , Glioma/genética , Polimorfismo de Nucleotídeo Único , Adulto , Alelos , Povo Asiático , Neoplasias Encefálicas/etnologia , Neoplasias Encefálicas/patologia , Estudos de Casos e Controles , Feminino , Expressão Gênica , Frequência do Gene , Genótipo , Glioma/etnologia , Glioma/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fatores de Risco , Análise de Sequência de DNARESUMO
Plant traits are important indices for regulating and controlling yield ability in soybean varieties. It is important to comprehensively study the quantitative trait locus (QTL) mapping for soybean plant traits, cloning related genes, and marker assistant breeding. In this study, 236 F2 generation plants and a derivative group were constructed by using Jiyu50 and Jinong18, obtained from Jilin Province. A total of 102 simple sequence repeat markers were used to construct a genetic linkage map. With 2 years of molecular and phenotypic data, QTL analyses and mapping were conducted for soybean maturity, plant height, main stem node, main stem branch, seed weight per plant, and more. Five main plant traits were analyzed via inclusive composite interval mapping using QTL IciMapping v2.2. Using one-dimensional scanning, a total of 30 QTLs were detected and distributed across 1 (A1), 4 (C2), and 12 (G). There were 9 linkage groups, including 16 major QTLs. Using two-dimensional scanning, 7 pairs of epistatic QTL interactions for maturity and plant height were detected in the soybean.
Assuntos
Mapeamento Cromossômico/métodos , Glycine max/genética , Locos de Características Quantitativas , Cromossomos de Plantas/genética , DNA de Plantas/análise , Ligação Genética , Hibridização Genética , Repetições de MicrossatélitesRESUMO
Seed number per pod is an important component of yield traits in soybean (Glycine max L.). In 2010, we identified a natural mutant with an increased number of four-seed pods from a soybean variety named 'Jinong 18' (JN18). Subsequent observations indicated that the trait was stably inherited. To identify and understand the function of genes associated with this mutant trait, we analyzed the genetic differences between the mutant (JN18MT01) and source variety (JN18) by transcriptome sequencing. Three types of tissues, axillary buds, unfertilized ovaries, and young pods at three different growth stages, V6, R1, and R3, were analyzed, respectively. The sequencing results yielded 55,582 expressed genes and 4183 differentially expressed genes (DEGs). Among these, the log2 ratio value of 162 DEGs was >10, and 13 DEGs had overlapping expression at three different growth stages. Comparisons of DEGs among three different growth stages yielded similar results in terms of the percentage of genes classified into each gene ontology (GO) category. DEGs were classified into 25 different functional groups in clusters of orthologous groups analysis. Proportions of the main functional genes differed significantly over developmental stages. A comparison of enriched pathways among the three developmental stages revealed that 646 unigenes were involved in 103 metabolic pathways. These results show that the development of four-seed pods is associated with a complex network involving multiple physiological and metabolic pathways. This study lays the foundation for further research on cloning and on the molecular regulation of genes related to the four-seed pod mutation.
Assuntos
Frutas/genética , Regulação da Expressão Gênica de Plantas , Glycine max/genética , Proteínas de Plantas/genética , Característica Quantitativa Herdável , Sementes/genética , Transcriptoma , Frutas/anatomia & histologia , Frutas/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Ontologia Genética , Redes Reguladoras de Genes , Sequenciamento de Nucleotídeos em Larga Escala , Redes e Vias Metabólicas/genética , Anotação de Sequência Molecular , Mutação , Fenótipo , Proteínas de Plantas/metabolismo , Sementes/crescimento & desenvolvimento , Glycine max/anatomia & histologia , Glycine max/crescimento & desenvolvimentoRESUMO
In order to investigate the genetic characteristics of soybean Leguminivora glycinivorella resistance and to improve soybean resistance insectivorous breeding efficiency by applying the multi-generation joint analysis method of the major gene plus polygene model, 5 pedigrees and generations (P1, F1, P2, F2, and F2:3) were used as the materials to perform the soybean L. glycinivorella resistance multi-generation joint analysis. The results showed that soybean resistance to L. glycinivorella was controlled and inherited by an additive major gene plus additive, dominant polygene. The major gene had a negative additive effect (d = -0.1633). The combination of the anti-L. glycinivorella genes showed negative heterosis. Because the polygene additive effects were positive, the polygene effects would increase the insect herbivory rate in the F1 generation. This hybrid combination showed an insect herbivory rate polygenic heritability of 21.9556 and 54.3490% in the F2 and F2:3 pedigrees, which presented a high heritability. Therefore, it was appropriate to perform the selective breeding of the insect herbivory rate in the late generation.
Assuntos
Genes de Plantas , Glycine max/genética , Lepidópteros/fisiologia , Modelos Genéticos , Herança Multifatorial , Animais , Cruzamento , Cruzamentos Genéticos , Glycine max/imunologia , Glycine max/parasitologiaRESUMO
Tuberous sclerosis complex (TSC) is an autosomal dominant neurogenetic disorder characterized by hamartomas in multiple organs and is caused by a wide spectrum of mutations in 1 of 2 causative genes (TSC1 or TSC2). Here, we present mutational analyses of the TSC1 and TSC2 genes in 4 cases of TSC in Chinese Han children, including 2 familial and 2 sporadic cases, using PCR and DNA sequencing of the entire coding region as well as exon-intron boundaries of these genes. Three mutations were identified in the TSC2 gene. Of these mutations, 2 mutations (c.3312-3313delGA and c.45delT) were novel, and the 3rd mutation (c.5238-5255del) was previously reported in Chinese Han and other populations. These mutations were not present in healthy family members or in 100 unrelated normal controls. The identification of these mutations in this study further expands the spectrum of known TSC2 gene mutations and contributes to prenatal molecular diagnosis and preimplantation genetic testing of TSC.
Assuntos
Povo Asiático/genética , Mutação , Esclerose Tuberosa/genética , Proteínas Supressoras de Tumor/genética , Adolescente , Angiofibroma/patologia , Sequência de Bases , Encéfalo/patologia , Pré-Escolar , China , Éxons , Feminino , Humanos , Lactente , Masculino , Linhagem , Análise de Sequência de DNA , Tomografia Computadorizada por Raios X , Esclerose Tuberosa/diagnóstico , Proteína 1 do Complexo Esclerose Tuberosa , Proteína 2 do Complexo Esclerose TuberosaRESUMO
We studied whether two IGF2 transcripts in common carp are similar to those found in zebrafish. The full-length IGF2a cDNA contains a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 1358 bp and an open reading frame of 612 bp, which encodes a 206-amino acid protein. A 6614-bp full-length IGF2a DNA molecule, including the 5'-flanking region, was isolated. Genomic DNA structure analysis revealed that the IGF2a gene contains four exons and three introns. Bioinformatics analysis indicated that the proteins encoded by IGF2a genes in common carp have one signal peptide and one apparent transmembrane region. Bootstrapping was performed 1000 times to obtain support values for each branch. The common carp IGF2a were clustered in one group, while the outgroup (common carp IGF1) clustered in another group. We identified two new single nucleotide polymorphisms in intron 2 of the gene. One polymorphism, A/N, can be found only in the Huanghe carp. The other polymorphism, C/N, can be found in both male Huanghe carp × female Heilongjiang carp and male Huanghe carp × female Jian carp. The second polymorphism, C/N, is primarily transferred from the male and may be related to heterosis.