RESUMEN
[This corrects the article DOI: 10.3892/etm.2017.4524.].
RESUMEN
Overexpression of pyrroline-5-carboxylate reductase 1 (PYCR1) has been associated with the development of certain cancers; however, no studies have specifically examined the role of PYCR1 in hepatocellular carcinoma (HCC). Based on The Cancer Genome Atlas expression array and meta-analysis conducted using the Gene Expression Omnibus database, we determined that PYCR1 was upregulated in HCC compared to adjacent nontumor tissues (P < 0.05). These data were verified using quantitative real-time polymerase chain reaction, western blotting, and immunohistochemistry analysis. Additionally, patients with low PYCR1 expression showed a higher overall survival rate than patients with high PYCR1 expression. Furthermore, PYCR1 overexpression was associated with the female sex, higher levels of alpha-fetoprotein, advanced clinical stages (III and IV), and a younger age (< 45 years old). Silencing of PYCR1 inhibited cell proliferation, invasive migration, epithelial-mesenchymal transition, and metastatic properties in HCC in vitro and in vivo. Using RNA sequencing and bioinformatics tools for data-dependent network analysis, we found binary relationships among PYCR1 and its interacting proteins in defined pathway modules. These findings indicated that PYCR1 played a multifunctional role in coordinating a variety of biological pathways involved in cell communication, cell proliferation and growth, cell migration, a mitogen-activated protein kinase cascade, ion binding, etc. The structural characteristics of key pathway components and PYCR1-interacting proteins were evaluated by molecular docking, and hotspot analysis showed that better affinities between PYCR1 and its interacting molecules were associated with the presence of arginine in the binding site. Finally, a candidate regulatory microRNA, miR-2355-5p, for PYCR1 mRNA was discovered in HCC. Overall, our study suggests that PYCR1 plays a vital role in HCC pathogenesis and may potentially serve as a molecular target for HCC treatment.
Asunto(s)
Carcinoma Hepatocelular/metabolismo , Transición Epitelial-Mesenquimal , Neoplasias Hepáticas/metabolismo , MicroARNs/metabolismo , Pirrolina Carboxilato Reductasas/metabolismo , Adulto , Animales , Carcinoma Hepatocelular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Hepáticas/genética , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , MicroARNs/genética , Persona de Mediana Edad , Simulación del Acoplamiento Molecular , Pirrolina Carboxilato Reductasas/genética , Ensayos Antitumor por Modelo de Xenoinjerto , delta-1-Pirrolina-5-Carboxilato ReductasaRESUMEN
Acute lung injury (ALI) is a life-threatening disorder with high rates of morbidity and mortality. Up to now, there are still no effective drugs for its therapies due to the complexity of its etiology and pathogenesis. In this present study, we investigated the protective effect of Nervilifordin F (NF) on ALI induced by intestinal ischemia/reperfusion (II/R) and its related mechanism. Firstly, the ALI model rats were induced through II/R, and treated with NF. Then, the pathological and cytokine level changes in the lung tissue of ALI rats were evaluated by hematoxylin and eosin and enzyme-linked immunosorbent assay (ELISA). The related genes expression level of mammalian target of rapamycin (mTOR) pathway and inflammasome were measured by real-time quantitative polymerase chain reaction (RT-qPCR), western blot and immunohistochemistry. Finally, the NF-protein complexes were predicted by SYBYL-X 2.0. The results indicated that NF can significant reduces the levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6 and IL-1ß, and inhibits the expression of inflammasome related genes (such as toll-like receptor 4 (TLR4), p65, NOD-like receptor protein 3 (NLRP3) and Caspase 1), thereby reduce inflammation in II/R-induced ALI rats. Moreover, NF can activate the expression of FK506 binding protein 25 (FKBP25) and down-regulate the expression of mTOR and p70 ribosomal protein S6 kinase 1 (p70S6K). In addition, molecular docking results showed that NF can be combined well with p70S6K, TLR4, mTOR and NLRP3, which further verified the inhibitory effect of NF on ALI inflammation. Therefore, the findings indicate that NF can alleviates II/R-induced inflammation of ALI rats by inhibiting inflammasome related genes and mTOR pathway, which expected to use as a potential drug for the treatment of ALI.
Asunto(s)
Lesión Pulmonar Aguda/prevención & control , Antiinflamatorios/farmacología , Inflamasomas/metabolismo , Enfermedades Intestinales/tratamiento farmacológico , Pulmón/efectos de los fármacos , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Daño por Reperfusión/tratamiento farmacológico , Serina-Treonina Quinasas TOR/metabolismo , Lesión Pulmonar Aguda/enzimología , Lesión Pulmonar Aguda/etiología , Lesión Pulmonar Aguda/patología , Animales , Citocinas/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Inflamasomas/genética , Enfermedades Intestinales/complicaciones , Enfermedades Intestinales/enzimología , Enfermedades Intestinales/patología , Pulmón/enzimología , Pulmón/patología , Masculino , Simulación del Acoplamiento Molecular , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Ratas Sprague-Dawley , Daño por Reperfusión/complicaciones , Daño por Reperfusión/enzimología , Daño por Reperfusión/patología , Transducción de Señal , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismoRESUMEN
Aberrant DNA replication is one of the driving forces behind oncogenesis. Furthermore, minichromosome maintenance complex component 3 (MCM3) serves an essential role in DNA replication. Therefore, in the present study, the diagnostic and prognostic value of MCM3 and its interacting proteins in hepatocellular carcinoma (HCC) were investigated. By utilizing The Cancer Genome Atlas (TCGA) database, global MCM3 mRNA levels were assessed in HCC and normal liver tissues. Its effects were further analyzed by reverse transcription-quantitative PCR (RT-qPCR), western blotting and immunohistochemistry in 78 paired HCC and adjacent tissues. Functional and pathway enrichment analyses were performed using the Search Tool for the Retrieval of Interacting Genes database. The expression levels of proteins that interact with MCM3 were also analyzed using the TCGA database and RT-qPCR. Finally, algorithms combining receiver operating characteristic (ROC) curves were constructed using binary logistic regression using the TCGA results. Increased MCM3 mRNA expression with high α-fetoprotein levels and advanced Edmondson-Steiner grade were found to be characteristic of HCC. Survival analysis revealed that high MCM3 expression was associated with poor outcomes in patients with HCC. In addition, MCM3 protein expression was associated with increased tumor invasion in HCC tissues. MCM3 and its interacting proteins were found to be primarily involved in DNA replication, cell cycle and a number of binding processes. Algorithms combining ROCs of MCM3 and its interacting proteins were found to have improved HCC diagnosis ability compared with MCM3 and other individual diagnostic markers. In conclusion, MCM3 appears to be a promising diagnostic biomarker for HCC. Additionally, the present study provides a basis for the multi-gene diagnosis of HCC using MCM3.
RESUMEN
BACKGROUND: Radiation sensitive 52 (RAD52) is an important protein that mediates DNA repair in tumors. However, little is known about the impact of RAD52 on hepatocellular carcinoma (HCC). We investigated the expression of RAD52 and its values in HCC. Some proteins that might be coordinated with RAD52 in HCC were also analyzed. METHODS: Global RAD52 mRNA levels in HCC were assessed using The Cancer Genome Atlas (TCGA) database. RAD52 expression was analyzed in 70 HCC tissues and adjacent tissues by quantitative real-time PCR (qRT-PCR), Western blotting and immunohistochemistry. The effect of over-expressed RAD52 in Huh7 HCC cells was investigated. The String database was then used to perform enrichment and functional analysis of RAD52 and its interactome. Cytoscape software was used to create a protein-protein interaction network. Molecular interaction studies with RAD52 and its interactome were performed using the molecular docking tools in Hex8.0.0. Finally, these DNA repair proteins, which interact with RAD52, were also analyzed using the TCGA dataset and were detected by qRT-PCR. Based on the TCGA database, algorithms combining ROC between RAD52 and RAD52 interactors were used to diagnose HCC by binary logistic regression. RESULTS: In TCGA, upregulated RAD52 related to gender was obtained in HCC. The area under the receiver operating characteristic curve (AUC) of RAD52 was 0.704. The results of overall survival (OS) and recurrence-free survival (RFS) indicated no difference in the prognosis between patients with high and low RAD52 gene expression. We validated that RAD52 expression was increased at the mRNA and protein levels in Chinese HCC tissues compared with adjacent tissues. Higher RAD52 was associated with older age, without correlation with other clinicopathological factors. In vitro, over-expressed RAD52 significantly promoted the proliferation and migration of Huh7 cells. Furthermore, RAD52 interactors (radiation sensitive 51, RAD51; X-ray repair cross complementing 6, XRCC6; Cofilin, CFL1) were also increased in HCC and participated in some biological processes with RAD52. Protein structure analysis showed that RAD52-RAD51 had the firmest binding structure with the lowest E-total energy (- 1120.5 kcal/mol) among the RAD52-RAD51, RAD52-CFL1, and RAD52-XRCC6 complexes. An algorithm combining ROC between RAD52 and its interactome indicated a greater specificity and sensitivity for HCC screening. CONCLUSIONS: Overall, our study suggested that RAD52 plays a vital role in HCC pathogenesis and serves as a potential molecular target for HCC diagnosis and treatment. This study's findings regarding the multigene prediction and diagnosis of HCC are valuable.
RESUMEN
The proliferation and migration of Schwann cells (SCs) are key events in the process of peripheral nerve repair. This is required to promote the growth of SCs and is a challenge during the treatment of peripheral nerve injury. Baicalin is a natural herb-derived flavonoid compound, which has been reported to possess neuroprotective effects on rats with permanent brain ischemia and neuronal differentiation of neural stem cells. The association of baicalin with neuroprotection leads to the suggestion that baicalin may exert effects on the growth of SCs. In the present study, the effects of baicalin on SCs of RSC96 were investigated. RSC96 SCs were treated with various concentrations of baicalin (0, 5, 10 or 20 µM) for 2, 4 and 6 days. Cell attachment, viability and gene expression were monitored via the MTT assay and reverse transcription-quantitative polymerase chain reaction. The gene expression levels of several neurotrophic factors, such as glial cell-derived neurotrophic factor, brain-derived neurotrophic factor and ciliary neurotrophic factor, which are considered important factors in the process of never cell regeneration, were detected. The results indicated that baicalin was able to promote the viability of RSC96 SCs in a dose-dependent manner and the concentration of 20 µM of baicalin exhibited the greatest cell viability and gene expression of the studied neurotrophic factors. The present findings suggested that baicalin likely affects SCs metabolism, through modulating the expression of neurotrophic factors. To conclude, the present study indicates that baicalin may be potential therapeutic agent for treating peripheral nerve regeneration.
RESUMEN
The proliferation and migration of Schwann cells (SCs) are essential in the process of peripheral nerve repair. A large amount of studies focused on the promotion of the growth of SCs for cell based therapy. Gastrodin (GAS), the main constituent of a Chinese traditional herbal medicine named Gastrodia elata Blume, has been reported to be associated with neuroprotective properties. Besides, GAS activated MAPK and PI3K signaling pathways which are often involved in growth of nerve cells were also reported. Based on the hypothesis that GAS may have an effect on SCs growth, we studied the effect of GAS on rat RSC96 Schwann cells (SCs) and further explored the underlying mechanism. Various concentration of GAS (0µM, 50µM, 100µM, and 200µM) was used for treatment of RSC96 SCs, with the cell proliferation and gene expression of several neurotrophic factors to be detected. Regulation of MAPK and PI3K signaling pathways were assayed by detecting phosphorylation of ERK1/2 and Akt. The results showed that GAS could effectively promote proliferation of RSC96 SCs in a dose- and time-dependent manner. The best performance was obtained at the concentration of 200µM. Exploration of the underlying mechanism showed that GAS probably affects SCs metabolism through inhibiting ERK1/2 phosphorylation and activating Akt phosphorylation in RSC96 SCs. This study may provide reference for its application in treatment of peripheral nerve injuries.
Asunto(s)
Alcoholes Bencílicos/farmacología , Proliferación Celular/efectos de los fármacos , Glucósidos/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/metabolismo , Células de Schwann/efectos de los fármacos , Células de Schwann/enzimología , Animales , Proliferación Celular/fisiología , Sistema de Señalización de MAP Quinasas/fisiología , Ratas , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiologíaRESUMEN
Synthesis of 1,3-propanediol (1,3-PD) from glycerol through the biotransformation process requires two steps, catalyzed by glycerol dehydratase (GDHt) and 1,3-PD oxidoreductase. GDHt is the rate-limiting enzyme in this process. All recombinant microorganisms for production of 1,3-PD so far utilized the natural genes that may not have been optimized. Two positions, which are 19.3A and 29.6A away from the active site in GDHt from Klebsiella pneumoniae, were subjected to saturation-mutagenesis and 38 mutants were characterized. The catalytic activity of a mutant in beta-subunit (beta-Q42F, 29.6A from the active site) was 8.3-fold higher than the wild type, and the enzyme efficiency of other two mutants beta-Q42L and beta-Q42S for substrate glycerol was 336-fold and 80-fold higher than that for 1,2-propanediol. This investigation supplied further evidence that distant mutations could be a good source of diversity and therefore, made a contribution to the toolbox of industrial enzyme improvement.