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Proteínas tirosina-fosfatase (PTPs) possuem papel fundamental na regulação da transdução de sinais e estão envolvidas em diversos processos fundamentais do ciclo celular. As Cdc25 (Cell Division Cycle 25) são fosfatases duais encontradas em todos os organismos eucarióticos e atuam em checkpoints do ciclo celular, permitindo ou inibindo o prosseguimento deste. Este grupo de proteínas pertence à classe de PTPs com atividade baseada em cisteína, apresenta domínio catalítico altamente conservado assim como o motivo catalítico, P-loop. Devido sua função, as Cdc25 são consideradas possíveis alvos terapêuticos para tratamento de câncer e sua interação com pequenas moléculas e inibidores tem sido investigada de forma que análises estruturais e de ligação das Cdc25 com inibidores podem elucidar aspectos importantes do mecanismo de ação destes além de direcionar para o desenho racional de fármacos. Interações cátion-π são interações intra ou intermoleculares não-covalentes que ocorrem entre uma espécie química catiônica, como o grupo guanidino de argininas, e uma das faces de um sistema π rico em elétrons, como dos anéis indólicos de triptofanos. Apesar de pouco discutidas na literatura, quando em comparação às interações não-covalentes mais convencionais, do ponto de vista energético as interações cátion-π são tão importantes na estruturação de proteínas quanto às ligações de hidrogênio ou pontes salinas. De fato estas interações são observadas com frequência em estruturas proteicas resolvidas. O domínio catalítico da Cdc25B possui diversas argininas expostas em sua superfície e um único resíduo de triptofano localizado na região C-terminal flexível, muito próximo do sítio catalítico da proteína. A flexibilidade de proteínas ou de regiões proteicas apresenta importante papel no reconhecimento entre biomoléculas participantes de vias de sinalização e tem sido muito estudada atualmente. Aqui, simulações de dinâmica molecular, experimentos de 1H-15N HSQC RMN, ensaios de cinética de inibição e de ancoragem molecular, evidenciam a existência de contatos cátion-π transientes na superfície de um importante membro da família das Cdc25, a Cdc25B, e de sítios de interação entre inibidores testados e a proteína com destaque a sítios na proximidades do P-loop, região próxima ao C-terminal desordenado, onde se demonstra estabilidade da interação com os pequenos ligantes
Protein tyrosine phosphatase (PTPs) play a fundamental role in the regulation of signal transduction and are involved in several fundamental processes of the cell cycle. Cdc25 (Cell Division Cycle 25) are dual phosphatases found in all eukaryotic organisms and act at checkpoints of the cell cycle, allowing or inhibiting its progression. This group of proteins belongs to the class of PTPs with cysteine-based activity, presenting a highly conserved catalytic domain as well as the catalytic motif, P-loop. Due to their function, Cdc25 are considered possible therapeutic targets for cancer treatment and their interaction with small molecules and inhibitors has been investigated so that structural and binding analyzes of Cdc25 with inhibitors can elucidate important aspects of their mechanism of action besides directing to rational drug design. Cation-π interactions are non-covalent intra- or intermolecular interactions that occur between a cationic chemical species, such as the guanidino group of arginines, and one of the faces of an electron-rich system, such as the indole rings of tryptophans. Although little discussed in the literature, when compared to more conventional non-covalent interactions, from the energetic point of view, cation-π interactions are as important in the structuring of proteins as hydrogen bonds or salt bridges. In fact, these interactions are frequently observed in solved protein structures. The catalytic domain of Cdc25B has several arginines exposed on its surface and a single tryptophan residue located in the flexible C-terminal region, very close to the catalytic site of the protein. The flexibility of proteins or protein regions plays an important role in the recognition between biomolecules participating in signaling pathways and has been extensively studied today. Here, molecular dynamics simulations, 1H-15N HSQC NMR experiments, inhibition kinetics and molecular anchoring assays, evidence the existence of transient cation-π contacts on the surface of an important member of the Cdc25 family, Cdc25B, and of sites of interaction between tested inhibitors and the protein, with emphasis on sites in the vicinity of the P-loop, a region close to the disordered C-terminus, where stability of the interaction with the small ligands is demonstrated
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cdc25 Phosphatases/analysis , Molecular Docking Simulation/methods , Molecular Dynamics Simulation/classificationABSTRACT
Aim To investigate the effects of dichloroacetate(DCA)combined with vitamin C(VC)on the malignant behavior of glioma U87 and U251 cells, and to explore the potential mechanism. Methods U87 and U251 cells were treated with different concentrations of DCA alone or in combination with 5 mmol·L-1 VC. The proliferation rate of each group was detected by CCK-8 method and the cooperative index was calculated. U87 and U251 cells were treated with DMSO, 15 mmol·L-1 DCA, 5 mmol·L-1 VC and their combination. The changes of clonal formation, reactive oxygen species content, apoptosis, cell cycle, migration and invasion were detected via in vitro experiments, while the proliferation of U251 cells in vivo in each group was detected by subcutaneous tumor-forming model. Western blot was used to detect the expression levels and degradation rates of BCL2A1 and CDC25A in each group of cells after network pharmacological analysis of DCA and VC targets and their value in glioma, and the expression levels of CDK4, CDK6, cytochrome C, caspase-7 and cleaved-caspase-7 were detected. Results The combined index of 15 mmol·L-1 DCA and 5 mmol·L-1 VC was the highest. Compared with the control and single drug groups, the clonal formation, migration and invasion ability of cells in combination group in vitro significantly decreased, the proliferation rate in vivo also decreased, and the content of reactive oxygen species, apoptosis rate and G1 phase arrest rate significantly increased. BCL2A1 and CDC25A proteins were important targets of DCA and VC in glioma. Compared with the control and single-drug groups, the expression levels of BCL2A1, CDC25A, CDK4, and CDK6 in the combination group were significantly reduced, and the expression levels of cytochrome C and cleaved-caspase-7 markedly increased, and the protein degradation rates of BCL2A1 and CDC25A significantly increased in the combination group. Conclusions VC can cooperate with DCA to promote the degradation of BCL2A1 and CDC25A, and inhibit the malignant behavior of glioma cells.
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Objective: Health information, including the labelling, must be presented in a manner that is easily understandable to consumers. In recent years, the European Union and the United States have introduced standards for providing health information in a way that is easy for consumers to understand. The Clear Communication Index (CCI) published by the Centers for Disease Control and Prevention (CDC) is a useful tool for this purpose. Unfortunately, there is no such tool in Japan. Therefore, focusing on the package labelling of foods with functional claims (FFCs) sold in Japan, we constructed a Functional Clear Communication Index (F-CCI) and evaluated the FFCs.Methods: The F-CCI was developed by six people, including university staff with pharmacist qualifications and public health experts, by referring to the CDC CCI. The evaluation of FFCs using the F-CCI was performed using the Delphi method, which is one of the formal consensus-building methods in the field of health and medical information. The evaluation was conducted by three qualified pharmacists on five FFC products, and the degree of internal agreement among the evaluators was calculated using Fleiss’ κ.Results: The F-CCI consisted of 18 items that assessed the FFC characteristics. After assessing the labelling of FFCs using the F-CCI, the scores of all the materials ranged between 70 and 80% on the F-CCI, and none achieved 90%, which was considered the acceptable standard (overall κ value_ 0.865). Moreover, it was clear that the ‘readability’ and ‘understandability’ of the labelling were inadequate.Conclusion: The F-CCI developed in this study for the objective evaluation of the labelling of FFC products will act as a tool that will subsequently lead to the proper understanding and use of FFCs by consumers. Further efforts are needed to build and disseminate such tools and user-friendly ways of providing relevant information.
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【Objective】 To explore critical regulatory genes in the hemoglobin switch process by analyzing transcriptomic data from the GSE6236, GSE17639 and GSE35102 datasets. 【Methods】 The mRNA expression profiles of the three datasets were downloaded from the GEO database and gene annotation was performed using the AnnoProbe package.The remove-BatchEffect function of the Limma package was used to remove batch effects. Weighted gene co-expression network analysis (WGCNA) was used to explore the most relevant modular genes in reticulocytes. The receiver operating characteristic curve (ROC) was used to assess the value of differential genes in differentiating between cord blood and adult peripheral blood reticulocytes. The GSE35102 dataset was used to validate changes in differential gene expression during hemoglobin transformation. Finally, real-time quantitative PCR was used to verify differential gene expression in cord blood and adult peripheral blood reticulocytes. 【Results】 Twelve genes showed differential expression in reticulocytes from cord blood and adult peripheral blood ( |logFC|≥1.5, P<0.05). WGCNA found that genes in the blue module were most strongly associated with reticulocytes (R2 =0.76,P<0.001). Of the five genes that overlapped between the two, only CDC42 showed differential expression in the combined dataset (t =3.776, P<0.001) and was able to better differentiate between reticulocytes in cord blood and adult peripheral blood. The expression of CDC42 varied significantly during the hemoglobin transformation process (Z = -2.908, P<0.01), and was significantly lower in adult reticulocytes compared to reticulocytes from cord blood (t =7.824, P <0.001). 【Conclusion】 The CDC42 gene is involved in the hemoglobin switching of reticulocytes and could be a potential therapeutic target for sickle cell disease.
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Introduction : Recent advances and improvements in technology in Neonatal Intensive Care Unit (NICU) over the past few decades have increased th survival of preterm infants. India ranks first amongst the number of preterm birth. Though the survival of preterm babies have increased Neuro-developmental morbidity amongst such babies still persist. Through this study we aim to establish a risk stratification tool and predict neurodevelopmental delay at 1 year of age. Materials and Methods : A total number of 77 babies were enrolled in the study after fulfilment of inclusion and exclusion criteria. Follow up till 1 year of age of corrected gestational age was done. Development assessment was done through Child Development Centre (CDC) grading, Amile-Tison Angle, Developmental Observation Card, Trivandrum Developmental Screening Chart and development quotient. Vision and Hearing Assessment was also done. Conclusion : Neuro-developmental outcome prediction at 1 year of age is inadequate and proper long term follow p is needed. Overall preterm babies needing extensive resuscitation and 5 minute APGAR <6 had much poor neurological outcome.
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Objective:To explore the value of cell division cycle 42 (CDC42) in the efficacy and prognosis evaluation of arterial infusion chemotherapy for pancreatic cancer.Methods:The clinical data of 100 patients with pancreatic cancer who underwent arterial infusion chemotherapy from January 2018 to January 2020 at Second People's Hospital of Wuhu were retrospectively analyzed, and all patients were divided into effective group (the complete remission and partial remission) and ineffective group (the stable disease and the progressive disease) according to the chemotherapy efficacy determined by CT. The clinicopathological characteristics of both groups were compared. The influencing factors of chemotherapy efficacy were determined by using multivariate logistic regression model analysis. The efficacy evaluated by CT examination was treated as the gold standard. The receiver operating characteristic (ROC) curve was used to analyze the value of CDC42 level predicting the efficacy of arterial infusion chemotherapy for pancreatic cancer patients before infusion chemotherapy. Survival analysis was performed by using Kaplan-Meier and log-rank test was also performed.Results:Among 100 patients with pancreatic cancer, there were 13 cases of complete remission, 30 cases of partial remission, 20 cases of stable disease, 37 cases of progressive disease; 43 cases in effective group and 57 cases in ineffective group. The proportions of tumor long diameter > 4 cm, TNM staging Ⅲ-Ⅳ, carcinoembryonic antigen (CA)199 > 37 U/ml, carcino-embryonic antigen (CEA) > 5 ng/ml, neutrophil-to-lymphocyte (NLR) > 2.8, serum total bilirubin > 34.2 μmol/L before infusion, CDC42 ≤ 1.11 μg/L, low differentiation degree and vascular invasion in ineffective group were higher than those in effective group (all P < 0.05). Tumor long diameter > 4 cm, TNM staging Ⅲ-Ⅳ, CA199 > 37 U/ml, CEA > 5 ng/ml before infusion, low differentiation degree, vascular invasion, and CDC42 ≤ 1.11 μg/L were independent risk factors for effectiveness of arterial infusion chemotherapy (all P < 0.05). The area under the ROC curve of CDC42 predicting the ineffectiveness of arterial infusion chemotherapy was 0.810 (95% CI 0.781-0.839, P <0.01), and the optimal cut-off value was 1.11 μg/L, the sensitivity was 96.25%, and the specificity was 63.13%. Survival curve analysis showed that the 2-year overall survival rate of patients with CDC42 > 1.11 μg/L was 58.93% which was greater than that of patients with CDC42 ≤ 1.11 μg/L (22.73%), and the difference was statistically significant ( χ2 = 14.99, P<0.001). Conclusion:CDC42 level is an independent influencing factor for the efficacy of arterial infusion chemotherapy in patients with pancreatic cancer, and it can effectively predict the prognosis of patients.
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Objective To investigate the effect of SP600125 on the proliferation, cell cycle, apoptosis and invasion of human cervical cancer HeLa cells. Methods CCK-8 method was used to detect the proliferation of HeLa cells treated with different concentrations of SP600125 at different time points. The 20 μmol/L of SP600125 was determined for subsequent experiments. Cell proliferation ability was detected using plate clone formation assay; nuclear morphology was observed by DAPI staining; cell cycle and apoptosis were measured by flow cytometry; cell migration and invasion were detected by cell scratch and Transwell methods; the mRNA and protein levels of p53, Mad2L1 and CDC20 were measured by qRT-PCR and Western blot after SP600125 treatment at different time points. Results Compared with control group (0.1%DMSO), cells proliferative activity were reduced by 10, 20, 30, 40 and 50 μmol/L SP600125 treatment for 24h. Compared with control group, the rate of apoptosis was significantly increased in SP600125 treatment groups, and the cell proportion in G2/M phase increased (P < 0.001), while the cell proportion in G0/G1 phases cells was reduced after SP600125 treatment for 24h and 48h (P < 0.001), and the clonal number, migration and invasion ability of HeLa cells also decreased significantly (P < 0.001). qRT-PCR and Western blot results showed a significant decrease in Mad2L1 mRNA and protein expression (P < 0.05) and a significant increase in p53 and CDC20 mRNA and protein expression (P < 0.01). Conclusion SP600125 can induce cell cycle arrest of cervical cancer HeLa cells in G2/M phase by upregulating p53 and CDC20 and downregulating Mad2L1 expression, and promote cell apoptosis to inhibit cell proliferation, migration and invasion.
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@#[Abstract] Objective: To analyze the expression of miR-185 and cell division cyclin 42 (CDC42) in osteosarcoma tissues and cells, and to preliminarily explore whether miR-185 affects the proliferation and migration of osteosarcoma MG63 cells by regulating CDC42. Methods: The cancer tissues and para-cancerous tissues of 28 patients with osteosarcoma that pathologically confirmed in the Fourth People's Hospital of Hengshui City from January 2020 to January 2021 were collected for this study. Immunohistochemistry was used to detect the expression of CDC42 in osteosarcoma tissues, and qPCR was used to detect the expression of miR-185 in osteosarcoma tissues. Dual-luciferase reporter gene experiment was applied to verify the targeting relationship between CDC42 and miR-185. According to different transfectants, MG63 cells were divided into miR-185 mimic group, miR-NC group, miR-185 inhibitor group, NC-inhibitor group, CDC42 group (transfected with CDC42 over-expression vector), and negative control (NC) group. The effects of miR-185 and CDC42 expression on the migration, proliferation and cell cycle of MG63 cells were detected by scratch healing assay, CCK-8 method and FCM, respectively. A nude mouse xenograft model was constructed by inoculating osteosarcoma MG63 cells. Immunohistochemistry, qPCR and WB methods were used to detect the effects of over-expression or knock-down of miR-185 on the expression of Ki67 and CDC42 in transplanted tumor tissues. Results: Compared with para-cancerous tissues, the expression of miR-185 in osteosarcoma tissues was significantly decreased, while the expression of CDC42 was significantly increased (all P<0.01). CDC42 was verified to be a target gene of miR-185. Compared with the control group, the migration and proliferation of MG63 cells in the miR-185 mimic group were inhibited (all P<0.01), while the migration and proliferation of MG63 cells in the CDC42 group were increased and the cell cycle was arrested in the S phase (all P<0.01). Compared with the miR-185 group, the migration and proliferation abilities of MG63 cells in the miR-185+CDC42 group were promoted, and the proportion of cells in S phase was increased (all P<0.01). Compared with the control group, the expression of Ki67 and CDC42 in the transplanted tumor tissues of miR-185 mimic group was significantly decreased (all P<0.01), while the opposite results were observed in miR-185 inhibitor group (all P<0.01). Conclusion: miR-185 is lowly expressed while CDC42 is highly expressed in osteosarcoma tissues. miR-185 can inhibit the proliferation and migration of osteosarcoma MG63 cells by negatively regulating the expression of CDC42.
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OBJECTIVE@#Osteosarcoma(OS) and Ewing's sarcoma (EWS) are the two most common primary malignant bone tumors in children. The aim of the study was to identify key genes in OS and EWS and investigate their potential pathways.@*METHODS@#Expression profiling (GSE16088 and GSE45544) were obtained from GEO DataSets. Differentially expressed genes were identified using GEO2R and key genes involved in the occurrence of both OS and EWS were selected using venn diagram. Gene ontology and pathway enrichment analyses were performed for the ensembl. Protein-protein interaction (PPI) networks were established by STRING. Further, UCSC was used to predict the transcription factors of the cell division cycke 5-like(CDC5L) gene, and GEPIA was used to analyze the correlation between the transcription factors and the CDC5L gene.@*RESULTS@#The results showed that CDC5L gene was the key gene involved in the pathogenesis of OS and EWS. The gene is mainly involved in mitosis, and is related to RNA metabolism, processing of capped intron-containing pre-mRNA, mRNA and pre-mRNA splicing.@*CONCLUSION@#CDC5L, as a key gene, plays a role in development of OS and EWS, which may be reliable targets for diagnosis and treatment of these primary malignant tumors.
Subject(s)
Child , Humans , Bone Neoplasms/pathology , Cell Cycle Proteins/genetics , Computational Biology , Gene Expression Profiling , Osteosarcoma/genetics , RNA-Binding Proteins/genetics , Sarcoma, Ewing/geneticsABSTRACT
The 90-kiloDalton (kD) heat shock protein (Hsp90) is a ubiquitous, ATP-dependent molecular chaperone whose primary function is to ensure the proper folding of several hundred client protein substrates. Because many of these clients are overexpressed or become mutated during cancer progression, Hsp90 inhibition has been pursued as a potential strategy for cancer as one can target multiple oncoproteins and signaling pathways simultaneously. The first discovered Hsp90 inhibitors, geldanamycin and radicicol, function by competitively binding to Hsp90's N-terminal binding site and inhibiting its ATPase activity. However, most of these N-terminal inhibitors exhibited detrimental activities during clinical evaluation due to induction of the pro-survival heat shock response as well as poor selectivity amongst the four isoforms. Consequently, alternative approaches to Hsp90 inhibition have been pursued and include C-terminal inhibition, isoform-selective inhibition, and the disruption of Hsp90 protein-protein interactions. Since the Hsp90 protein folding cycle requires the assembly of Hsp90 into a large heteroprotein complex, along with various co-chaperones and immunophilins, the development of small molecules that prevent assembly of the complex offers an alternative method of Hsp90 inhibition.
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Objective:To analyze the expression of cell differentiation cyclin 45 (CDC45) in hepatocellular carcinoma (HCC) tissues, to explore the relationship between CDC45 and the prognosis of HCC, and to explore the possibility of CDC45 as a potential biomarker and therapeutic target for HCC.Methods:Bioinformatics methods were used to analyze the mRNA levels of CDC45 in HCC tissues and normal tissues, and to analyze the relationship between its expression and the survival rate of HCC patients. The clinicopathological data of 90 patients with HCC undergoing surgical treatment were retrospectively analyzed. Immunohistochemical method was used to detect the expression level of CDC45 protein in HCC tissue and normal tissues, and to analyze its relationship with clinicopathological characteristics of patients with HCC.Results:CDC45 is highly expressed in colorectal cancer, breast cancer, prostate cancer, lung cancer, liver cancer, etc. The expression of CDC45 in HCC tissues was higher than that in normal tissues adjacent to cancer ( P<0.05). The expression of CDC45 is significantly related to the clinical prognosis of HCC patients ( P<0.001), and the higher the expression of CDC45, the lower the disease-free survival rate and overall survival rate of HCC patients. CDC45 level is significantly correlated with the number of tumor nodules ( P=0.003) and tumor size ( P=0.003) in HCC patients, but the correlation between CDC45 level and age, gender and alpha-fetoprotein (AFP) levels is not statistically significant (all P>0.05). Conclusions:CDC45 plays an important role in the progression of HCC and is expected to become a therapeutic target and potential biomarker for HCC.
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Objective:The aim of this article is to analyze the epidemiology of COVID-19, comparison of routes of transmission in children and adults, comparing the clinical symptoms in adults and children, treatment protocol to be followed and possible treatment options duringthis pandemic. Data Sources:Data is collected from Pubmed, Medline and Embase databases. Discussion from Previous References Used: Few studies have been done to analyze its effect on children comparing the symptoms of adults and children. Also very lesswork is done to analyze the special precautions taken while doing dental treatment during this pandemic. Conclusion:The widespread effect of Coronavirus (COVID-19) or SARS-CoV-2 has created health concerns in the world. Although efforts have been taken to control the disease, it is still increasing due to the community spread. Health professionals may get patients in their practice with this infection and should prevent its spread.Clinical Significance of this Article:It will help us to differentiate the specific clinical symptoms of adult and pediatric patients coming to dental clinics and the special considerations for them including the emergency dental treatment during COVID-19
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Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with a primary target on the human respiratory system. Coronavirus disease was first discovered in Wuhan, China in December 2019 and has currently become a global pandemic. A lot is still unknown about COVID -19 pathogenesis. Prompt assessment, adequate follow up, test and retest of recovered cases to corroborate immune related considerations will go a long way to understand the pathogenesis
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A series of 2,3-dioxoindolin-N-phenylacetamide derivatives was evaluated for inhibitory activity against CDC25B and PTP1B enzymes. Most of the derivatives showed inhibitory activity against CDC25B (IC50 = 3.2-23.2 µg/mL) and PTP1B (IC50 = 2.9-21.4 µg/mL). Compound 2h showed the most inhibitory activity in vitro with IC50 values of 3.2 and 2.9 µg/mL against CDC25B and PTP1B, respectively, compared with the reference drugs Na3VO4 (IC50 = 2.7 µg/mL) and oleanolic acid (IC50 = 2.3 µg/mL). The results of selectivity experiments showed that the 2,3-dioxoindolin-N-phenylacetamide derivatives were selective inhibitors against CDC25B and PTP1B. Enzyme kinetic experiments demonstrated that compound 2h was a specific inhibitor with the typical characteristics of a mixed inhibitor. In cytotoxic activity assays compound 2h had potent activity against A549, HeLa, and HCT116 cell lines. In addition, compound 2h showed potent tumor inhibitory activity in a colo205 xenograft model in vivo.
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ACK1 (activated Cdc42-associated kinase) is a non-receptor tyrosine kinase, originally identified by its binding to the GTP-binding small GTPase Cdc42. It is widely expressed in human tissues and activated by various extracellular growth factors such as EGF, PDGF and TGF-β. The activated ACK1 mediates the signaling cascade by interacting with downstream effectors followed by their phosphorylation. In recent years, researchers have investigated the biological functions of ACK1 and its roles in cancer research. The gene amplification and overexpression of ACK1 is associated with a poor prognosis and metastasis in a variety of cancers including lung, ovarian and prostate cancers. Therefore, the development of small molecule inhibitors of ACK1 provides promising opportunities for cancer-targeted therapy. In this review, we briefly describe recent advances in understanding the activation and biological function of ACK1 and introduce its novel inhibitors with potential therapeutic activities in preclinical studies.
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Objective: To study the effect of rocaglaol from Aglaia odorata on HepG2 proliferation and to explore the potential anti-tumor mechanism. Methods: The MTT, colony formation, EdU incorporation, and CFDA-SE assays were used to determine the anti-proliferative activity of rocaglaol in HepG2 cells. Apoptosis and cell cycle distribution effect induced by rocaglaol were carried out by flow cytometry. The effect of rocaglaol on protein involved in the G2/M checkpoint and the MAPK pathway were performed by Western blotting analysis. Results: Rocaglaol significantly inhibited the viability of HepG2 cells in a dose-dependent and time-dependent manner. Rocaglaol was more effective than doxorubicin in the growth inhibition of HepG2 cells. However, rocaglaol-induced cytotoxicity in normal human hepatic cell line L02 was lower than that of doxorubicin. Treatment with different concentrations of rocaglaol at 48 h caused G2/M cell cycle progression inhibition, rather than apoptosis in HepG2 cells. Rocaglaol can significantly reduce the expression of G2/M cell cycle-regulating proteins cdc25C, cdc2, and cyclin B1 as well as increase the expression of ERK and JNK phosphorylation levels. Further study found that U0126 can partly abrogate the anti-proliferative activity in HepG2 cells, G2/M phase arrest and the reduction in the protein expression levels of cdc2 and cdc25C induced by rocaglaol. Conclusion: Our results demonstrated that rocaglaol was superior to doxorubicin in the inhibition of HepG2 cells proliferation and the selectivity of L02 cell activity. We provided evidence that the rocaglaol had the ability to continuously over-activate the ERK signaling in HepG2 cells, leading to the inhibition of cell proliferation through G2/M phase arrest.
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OBJECTIVE@#To investigate the expression of CDC25A in non- small cell lung cancer (NSCLC) tissues and explore its correlation with the clinicpathological features of the patients and the expressions of let-7a1 and let-7c.@*METHODS@#We collected surgical specimens of pathologically confirmed NSCLC tissues and paired adjacent lung tissues from 44 patients and tissues of benign lung lesions from 9 patients. The expressions of CDC25A protein and mRNA in the tissues were detected by immunohistochemistry and fluorescence quantitative RT-PCR, respectively; the expressions of let-7a1 and let-7c mRNA were detected using tail-adding fluorescence quantitative RT-PCR.@*RESULTS@#The positivity rate of CDC25A protein expression was significantly higher in NSCLC tissues than in the adjacent tissues and benign pulmonary lesions (@*CONCLUSIONS@#The expression level of CDC25A is significantly increased in NSCLC with a negative correlation with Let-7c expression, which identifies CDC25A as a possible downstream target gene of Let-7c.
Subject(s)
Humans , Carcinoma, Non-Small-Cell Lung/genetics , Gene Expression Regulation, Neoplastic , Lung , Lung Neoplasms/genetics , Lymphatic Metastasis , MicroRNAs , RNA, Messenger/genetics , cdc25 PhosphatasesABSTRACT
@#Objective: To explore the genes that may be regulated by cell division cycle 25A (CDC25A) with gene chip technology, and to elucidate and verify that CDC25A has a regulatory effect on the expression of liver cancer related genes. Methods: CDC25A expression in human liver cancer HepG2 cells was silenced by siRNA interference technology and a nude mouse xenograft model of liver cancer was successfully constructed in our previous research. Affymetrix human gene expression profiling microarray was used to further screen differentially expressed genes (DEGs) after silencing CDC25A in liver cancer xenografts, and GO analysis and KEGG analysis were performed. Some of the DEGs were verified by qPCR. Results: The chip screened 188 DEGs in liver cancer xenograft tissues after CDC25A silence, including 78 up-regulated genes and 110 down-regulated genes. These DEGs mainly involved in cell proliferation, apoptosis, protein complex binding, extracellular space, etc., and associated with the changes in pathways such as focal adhesions and extracellular matrix (ECM) receptor interactions. qPCR showed that the expression of HIPK2 mRNA was up-regulated and the mRNA expressions of (microfibrillar-associated protein 5(MFAP5) and cyclin D1 (CCND1) were down-regulated, which were consistent with the results of microarray detection. Conclusion: Using human gene expression profiling chip, the DEGs in liver cancer xenograft tissues in nude mice after silencing CDC25Awere successfully screened, providing effective clues for exploring the effect of CDC25Aon the growth of liver cancer.
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@#AIM: To observe the effect and mechanism of MicroRNA-34a on senescence and apoptosis of human lens epithelial cell line SRA01/04.<p>METHODS: MicroRNA-34a expression levels in ARC lens and transparent lens epithelial cells were detected by qRT-PCR. MicroRNA-34a mimics, MicroRNA-34a inhibitors and empty liposome(control group)were transfected into SRA01/04 cells by liposome transfection kit. Annexin V-FITC/PI double staining was used to detect the effect of MicroRNA-34a on the apoptosis of human lens cell line SRA01/04. The expression of Cdc42 and Rac1 protein was detected by western blot. <p>RESULTS: The expression level of MicroRNA-34a in anterior capsular tissue of transparent lens was significantly lower than that in ARC anterior capsular tissue(<i>P</i><0.05). The positive rates of SA-β-gal in the MicroRNA-34a mimics group, the control group and the MicroRNA-34a inhibitors group were(87.56±2.34)%,(12.22±2.74)% and(3.45±0.45)%, respectively. The positive rates of SA-β-gal in the MicroRNA-34a mimics group was significantly higher than the control group, while the SA-β-gal positive rate in the MicroRNA-34a inhibitors group was significantly lower than that in the control group(<i>P</i><0.05). The apoptosis rate of the MicroRNA-34a inhibitors group, control group and MicroRNA-34a mimics group were(5.87±1.22)%,(12.26±2.14)% and(29.45±3.12)%, respectively. The apoptosis rate of the MicroRNA-34a mimics group was significantly higher than that of the control group, while that of the MicroRNA-34a inhibitors group was significantly lower than that of the control group(<i>P</i><0.05). The expressions of Cdc42 and Rac1 in the MicroRNA-34a mimics group were significantly higher than those in the control group(<i>P</i><0.05), while the expressions of Cdc42 and Rac1 in the MicroRNA-34a inhibitors group were significantly lower than those in the control group(<i>P</i><0.05).<p>CONCLUSION: MicroRNA-34a may promote the senescence and apoptosis of human lens epithelial cells by up-regulating Cdc42 and Rac1.
ABSTRACT
Abstract Introduction: Cell division cycle-7 protein is a serine/threonine kinase that has a basic role in cell cycle regulation and is a potential prognostic or therapeutic target in some human cancers. Objectives: This study investigated the expression of cell division cycle-7 protein in benign and malignant salivary gland tumors and also its correlation with clinicopathologic factors. Methods: Immunohistochemical expression of cell division cycle-7 was evaluated in 46 cases, including 15 adenoid cystic carcinoma, 12 mucoepidermoid carcinoma, 14 pleomorphic adenoma, and 5 normal salivary glands. Cell division cycle-7 expression rate and intensity were compared statistically. Results: The protein was expressed in almost all tumors. The intensity and mean of cell division cycle-7 expression were higher in malignant tumors in comparison with pleomorphic adenomas (p = 0.000). The protein expression was correlated with tumor grades (p = 0.000). Conclusions: The present study demonstrated cell division cycle-7 overexpression in malignant salivary gland tumors in comparison with pleomorphic adenomas, and also a correlation with tumor differentiation. Therefore, this protein might be a potential prognostic and therapeutic target for salivary gland tumors.
Resumo Introdução: A cell division cycle-7 é uma serina/treonina quinase que tem um papel básico na regulação do ciclo celular e é um potencial marcador prognóstico ou terapêutico em alguns tipos de câncer humano. Objetivos: Este estudo investigou a expressão de cell division cycle-7 em tumores de glândulas salivares benignos e malignos e também sua correlação com fatores clínico-patológicos. Método: A expressão imuno-histoquímica de cell division cycle-7 foi avaliada em 46 casos, incluindo 15 carcinomas adenoide císticos, 12 carcinomas mucoepidermoides, 14 adenomas pleomórficos e 5 glândulas salivares normais. A taxa de expressão e a intensidade da proteína cell division cycle-7 foram comparadas estatisticamente. Resultados: A proteína foi expressa em quase todos os tumores. A intensidade e a média da expressão de cell division cycle-7 foram maiores em tumores malignos em comparação com adenoma pleomórfico (p = 0,000). A expressão da proteína foi correlacionada com os graus do tumor (p = 0,000). Conclusões: O presente estudo demonstrou a superexpressão de cell division cycle-7 em tumores malignos de glândulas salivares quando comparada com o adenoma pleomórfico, além de uma correlação com a diferenciação de tumores. Portanto, essa proteína pode ser um potencial marcador prognóstico e terapêutico para tumores de glândulas salivares.