RESUMO
OBJECTIVE: To explore the effect of intervention of E-cadherin (E-cad) and B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1) mediated by transcription activator-like effector nuclease (TALEN) on the biological behaviors of nasopharyngeal carcinoma cells.â© Methods: Multi-locus gene targeting vectors pUC-DS1-CMV-E-cad-2A-Neo-DS2 and pUC-DS1-Bmi-1 shRNA-Zeo-DS2 were constructed, and the E-cad and Bmi-1 targeting vectors were transferred with TALEN plasmids to CNE-2 cells individually or simultaneously. The integration of target genes were detected by PCR, the expressions of E-cad and Bmi-1 were detected by Western blot. The changes of cell proliferation were detected by cell counting kit-8 (CCK-8) assay. The cell cycle and apoptosis were detected by flow cytometry. The cell migration and invasion were detected by Transwell assay.â© Results: The E-cad and Bmi-1 shRNA expression elements were successfully integrated into the genome of CNE-2 cells, the protein expression level of E-cad was up-regulated, and the protein expression level of Bmi-1 was down-regulated. The intervention of E-cad and Bmi-1 didn't affect the proliferation, cell cycle and apoptosis of CNE-2 cells, but it significantly inhibited the migration and invasion ability of CNE-2 cells. Furthermore, the intervention of E-cad and Bmi-1 together significantly inhibited the migration ability of nasopharyngeal carcinoma cells compared with the intervention of E-cad or Bmi-1 alone (all P<0.01).â© Conclusion: The joint intervention of E-cad and Bmi-1 mediated by TALEN can effectively inhibit the migration and invasion of nasopharyngeal carcinoma cells in vitro, which may lay the preliminary experimental basis for gene therapy of human cancer.
Assuntos
Apoptose/fisiologia , Caderinas/fisiologia , Carcinoma/patologia , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Neoplasias Nasofaríngeas/patologia , Complexo Repressor Polycomb 1/fisiologia , Proteínas Proto-Oncogênicas/fisiologia , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição/metabolismo , Animais , Caderinas/genética , Carcinoma/genética , Carcinoma/metabolismo , Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Vetores Genéticos , Humanos , Técnicas In Vitro , Camundongos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/metabolismo , Complexo Repressor Polycomb 1/genética , Proteínas Proto-Oncogênicas/genéticaRESUMO
The complete mitochondrial genome of Paracheirodon innesi was determined using the next-generation sequencing (NGS). The mitochondrial genome was 16 962 bp long and consisted of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. The gene arrangement and composition of P. Innesi mitochondrial genome was similar to that of most other vertebrates. Phylogenetic tree was constructed based on the complete mitogenomes of the species and closely related 13 teleost species to assess their phylogenetic relationship and evolution.
Assuntos
Characidae/genética , Genes Mitocondriais , Genoma Mitocondrial , Filogenia , Animais , Composição de Bases , Sequência de Bases , DNA Mitocondrial , Ordem dos Genes , Tamanho do Genoma , Sequenciamento de Nucleotídeos em Larga Escala , Análise de Sequência de DNARESUMO
The palladium-catalyzed enantioselective intramolecular C-H arylation of N-(2-haloaryl)-P,P-diphenylphosphinic amides furnishes P-stereogenic phosphine oxide derivatives in 61-99% yield with 88-97% ee. The catalyst generated in situ from a TADDOL-derived phosphoramide ligand and Pd(dba)2 is optimum in terms of yield and enantioselectivities.
