RESUMO
This study investigated the efficacy and feasibility of inducing the differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) into hepatocyte-like cells in vitro using Sprague Dawley rats, as a model of hepatocyte generation for cell transplantation. BMSCs were isolated and grown using the adherent method and exposed to 5 or 10% liver tissue homogenate, before being collected for analysis after 0, 7, 14, and 21 days. Immunofluorescence and western blotting were employed to detect the liver-specific markers a-fetoprotein (AFP) and albumin (ALB). Supernatant urea content was also measured to verify that differentiation had been induced. After 7 days in the presence of 10% liver tissue homogenate, BMSCs demonstrated hepatocyte-like morphological characteristics, and with prolonged culture time, liver-specific markers were gradually produced at levels indicating cell maturation. AFP expression peaked at 14 days then began to decrease, while both urea and ALB levels increased with induction time. Overall, marker expression in the 5% homogenate group was less than or equal to the 10% group at each time point. Thus, in a rat model, liver tissue homogenate obtained from partial hepatectomy can induce the differentiation of BMSCs into hepatocyte-like cells. This method is simple, feasible, and has remarkable real-world application potential.
Assuntos
Diferenciação Celular , Hepatócitos/citologia , Fígado/metabolismo , Células-Tronco Mesenquimais/citologia , Albuminas/genética , Albuminas/metabolismo , Animais , Células Cultivadas , Proteínas Fetais/genética , Proteínas Fetais/metabolismo , Hepatócitos/metabolismo , Fígado/química , Masculino , Células-Tronco Mesenquimais/metabolismo , Ratos , Ratos Sprague-Dawley , Frações Subcelulares/química , Frações Subcelulares/metabolismoRESUMO
It has been reported that the estrogen receptor alpha (ESR1) rs9340799 polymorphism is associated with age at menarche (AAM). However, recent investigations have generated inconsistent results. This study aimed to establish a more precise estimation of the association between this polymorphism and AAM. A meta-analysis was conducted based on an in silico literature search using PubMed. Six studies presenting continuous data, including ESR1 rs9340799 genotype frequencies, were selected. Effect size was estimated using Hedges' adjusted g with 95% confidence intervals (CIs), which were calculated based on the standardized mean difference between groups of subjects and different genotypes. No evidence of an association between the ESR1 rs9340799 polymorphism and AAM was found in the pooled continuous data under any genotype comparison (AA vs GG+AG: Hedges' g = -0.085, 95%CI = -0.202-0.032, P = 0.156; GG vs AA+AG: Hedges' g = 0.143, 95%CI = -0.041-0.327, P = 0.129; A vs G: Hedges' g = 0.187, 95%CI = -0.032-0.406, P = 0.095). Moreover, a funnel plot generated using this data was found to be symmetrical using the Egger (P = 0.797) and Begg tests (P = 0.851), indicating the absence of publication bias. In summary, our meta-analysis shows that the ESR1 rs9340799 polymorphism is not a significant, independent contributing factor to AAM. To validate this finding, further studies involving larger numbers of participants are needed.
Assuntos
Receptor alfa de Estrogênio/genética , Menarca/genética , Polimorfismo de Nucleotídeo Único , Feminino , Genótipo , HumanosRESUMO
Mitochondria are closely associated with cell survival, and it is of interest to determine whether apoptosis pathways, which are mediated by mitochondria, are involved in liver regeneration (LR). To identify the mechanisms underlying mitochondria-mediated apoptosis during rat LR, we used the Rat Genome 230 2.0 Array to investigate changes in gene expression. Next, we searched the GO and NCBI databases for genes associated with apoptosis mediated by mitochondria, and QIAGEN and KEGG databases for any related signaling pathways. The expression profile function (Et) was then used to calculate the activity level of known signaling pathways associated with apoptosis. The results revealed the expression of 436 genes associated with apoptosis signaling pathways, among which 152 were confirmed to be primarily related to LR. Overall, 99, 136, 95, and 91 genes were first expressed during the initiation [0.5-4 h after partial hepatectomy (PH)], G0/G1 transition (4-6 h after PH), cell proliferation (6-66 h after PH), and redifferentiation and structural reconstruction (66-144 h after PH) phases, demonstrating that LR-related genes were primarily induced in the initiation phase, and were then expressed across multiple phases. Analysis using the gene synergy formula (Et) showed that caspase-dependent and DNA fragment-related/unrelated pathways induced apoptosis in the early and late periods of LR, and the caspase-independent and DNA fragment-related/unrelated pathways almost in the whole process. Therefore, these results show that several apoptosis pathways regulate LR in rat.
Assuntos
Apoptose , Regeneração Hepática , Mitocôndrias/genética , Transcriptoma , Animais , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Proliferação de Células , Perfilação da Expressão Gênica , Masculino , Ratos Sprague-Dawley , Transdução de SinaisRESUMO
The death receptor and endoplasmic reticulum (ER) are closely related to cell apoptosis, and it is worth studying whether the apoptosis pathways mediated by them are involved in liver regeneration. To understand the mechanism underlying death receptor- and ER-mediated apoptosis during rat liver regeneration, we used the Rat Genome 230 2.0 Array to determine the changes in gene expression. We then searched the gene ontology (GO) and NCBI databases for genes associated with cell apoptosis mediated by the death receptor and ER. QIAGEN and KEGG databases were used for the related signaling pathways. We used the expression profile function to calculate the activity levels of the known apoptosis signaling pathways. The results of our study showed that the initial gene expression numbers in initiation, G0/G1 transition, cell proliferation, and redifferentiation and structural reconstruction phases were 32, 25, 44, and 29, respectively. This demonstrates that liver regeneration-related genes primarily start their expression in the initiation phase and work differently in each phase. By calculation and analysis using the gene synergy formula, it was suggested that the apoptosis signaling pathways [FAS, death receptor 3 (DR3), tumor necrosis factor receptor 1 (TNFR1), and ER] induced cell apoptosis in whole liver regeneration and anti-apoptosis pathways (DR3 and TNFR2) restrained apoptosis in the early phase of liver regeneration. In summary, these apoptosis pathways coordinated and regulated quality and quantity of the regenerating liver cells.
Assuntos
Apoptose , Retículo Endoplasmático/metabolismo , Regeneração Hepática , Receptores de Morte Celular/genética , Animais , Proliferação de Células , Pontos de Checagem da Fase G1 do Ciclo Celular , Fígado/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Receptores de Morte Celular/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Transdução de SinaisRESUMO
Chromosome instability is a common feature of tumor cells, and may be an important mechanism in tumor formation. Nuclear division cycle 80 (NDC80) is closely associated with the stability of chromosomes. Therefore, we investigated the relationship between NDC80 and development of colon cancer using a range of methods. Western blotting and immunohistochemistry were employed to determine the expression of this protein in different colon cells and tissues, cell proliferation was measured with an MTT assay, levels of proliferating cell nuclear antigen were examined by immunofluorescence, and cell migration was observed using wound healing tests. Our results showed that the expression of NDC80 in colon cancer cells (CACO2, HCT8, HCT116, and SW480) and tissues (from 20 patients) was higher than that in controls. Moreover, cell proliferation and migration rates were elevated in cells transfected with NDC80 compared to control groups. In summary, NDC80 promotes the proliferation and metastasis of colon cancer cells, and may constitute a new target for gene therapy in treating this disease. Combined with clinicopathological grading, measurement of positive NDC80 expression may be helpful in diagnosing and estimating the prognosis of colon cancer patients.
Assuntos
Adenocarcinoma/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias do Colo/metabolismo , Proteínas Nucleares/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Adulto , Biomarcadores Tumorais/genética , Células CACO-2 , Movimento Celular , Proliferação de Células , Instabilidade Cromossômica , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Proteínas do Citoesqueleto , Feminino , Células HCT116 , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Proteínas Nucleares/genéticaRESUMO
The liver is the human body largest digestive and metabolic organ, and a very important immune organ. This paper discusses the location and morphology of hepatic sinusoidal endothelial cells, dendritic cells, hepatic stellate cells, and Kupffer cells in the liver and their role in regulating immune functions. Therefore, here we provide a preliminary understanding of the immune regulatory function of liver cells, and information on the occurrence and treatment of liver diseases.