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Resumen Introducción: Las células dendríticas (CD) están involucradas en el reconocimiento, respuesta y modulación inmunológicos relacionados con la aparición del cáncer. Objetivo: Explorar el mecanismo de las CD en la inhibición de la autofagia de las células del hepatoma. Métodos: Células mononucleares de sangre periférica humana se aislaron mediante centrifugación en gradiente de densidad de Ficoll y se indujeron en CD, las cuales fueron cocultivadas con células HepG2 por ensayo de migración Transwell. La actividad de las células HepG2 se determinó mediante ensayo CCK8. La expresión del índice de autofagia LC3 se midió con análisis de transferencia Western y la expresión y secreción de citocinas mediante qRT-PCR y ELISA. Resultados: En el sistema de cocultivo, las CD redujeron la viabilidad de HepG2; la expresión de IL-2, IL-12, IL-10 e IFN-γ en CD también se inhibió significativamente, si bien IL-2 e IFN-γ aún se expresaron 0.6 y 0.53 más que en el grupo de control. Conclusión: Las CD pueden regular la autofagia de las células del carcinoma hepatocelular. El mecanismo puede estar relacionado con la síntesis y liberación de citocinas como IL-2, IL-12 e IFN-γ por parte de las CD.
Abstract Introduction: Dendritic cells (DC) are involved in immune recognition, response and immunomodulation mechanisms related to the onset of cancer. Objective: To explore DCs mechanism in the inhibition of autophagy in hepatoma cells. Methods: Human peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation and induced into DCs, which were co-cultured with HepG2 cells by Transwell migration assay. HepG2 cell activity was determined using the CCK8 assay. LC3 autophagy index expression was measured with Western blot analysis, and the expression and secretion of cytokines, with qRT-PCR and ELISA. Results: In the co-culture system, DCs were able to reduce HepG2 cells viability; IL-2, IL-12, IL-10 and IFN-γ expression in DCs was also significantly inhibited, although IL-2 and IFN-γ were still expressed 0.6 and 0.53 more than in the control group. Conclusion: DCs can regulate autophagy in hepatocellular carcinoma cells. The mechanism may be related to the synthesis and release of cytokines such as IL-2, IL-12 and IFN-γ by DCs.
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Objective: The objective of this paper was to investigate hub genes of postmenopausal osteoporosis (PO) utilizing benchmarked dataset and gene regulatory network (GRN). Materials and Methods: To achieve this goal, the first step was to benchmark the dataset downloaded from the ArrayExpress database by adding local noise and global noise. Second, differentially expressed genes (DEGs) between PO and normal controls were identified using the Linear Models for Microarray Data package based on benchmarked dataset. Third, five kinds of GRN inference methods, which comprised Zscore, GeneNet, context likelihood of relatedness (CLR) algorithm, Partial Correlation coefficient with Information Theory (PCIT), and GEne Network Inference with Ensemble of trees (Genie3), were described and evaluated by receiver operating characteristic (ROC) and precision and recall (PR) curves. Finally, GRN constructed according to the method with best performance was implemented to conduct topological centrality (closeness) for the purpose of investigate hub genes of PO. Results:A total of 236 DEGs were obtained based on benchmarked dataset of 20,554 genes. By assessing Zscore, GeneNet, CLR, PCIT, and Genie3 on the basis of ROC and PR curves, Genie3 had a clear advantage than others and was applied to construct the GRN which was composed of 236 nodes and 27,730 edges. Closeness centrality analysis of GRN was carried out, and we identified 14 hub genes (such as TTN, ACTA1, and MYBPC1) for PO. Conclusion: In conclusion, we have identified 14 hub genes (such as TN, ACTA1, and MYBPC1) based on benchmarked dataset and GRN. These genes might be potential biomarkers and give insights for diagnose and treatment of PO