المؤلفون: Shao-Hsuan Kao, Jiun-Tsai Lin, Wei-Wen Su, Han-Min Chen, Jen-Yu Huang

المصدر: International Journal of Medical Sciences

مصطلحات موضوعية: AMPK, p53, Carcinoma, Hepatocellular, Cyclin A, Drug Evaluation, Preclinical, AMP-Activated Protein Kinases, 03 medical and health sciences, 0302 clinical medicine, Cyclin-dependent kinase, Humans, Protein kinase A, adenine, hepatocellular cell, biology, p21, Kinase, Chemistry, Cell growth, Cell Cycle, Liver Neoplasms, apoptosis, General Medicine, Hep G2 Cells, Cell cycle, digestive system diseases, Cell culture, Bax, biology.protein, Cancer research, 030211 gastroenterology & hepatology, Research Paper

الوصف: Background: Adenine exhibits potential anticancer activity against several types of malignancies. However, whether adenine has anticancer effects on hepatocellular carcinoma (HCC) cells is incompletely explored. Methods: Human HCC cell lines HepG2 and SK-Hep-1 (p53-wild type) and Hep3B (p53-deficient) were used as cell model. Cell growth and cell cycle distribution were determined using MTT assay and flow cytometric analysis, respectively. Protein expression and phosphorylation were assessed by Western blot. Involvement of AMP-activated protein kinase (AMPK) was evaluated using specific inhibitor and small inhibitory RNA (siRNA). Results: Adenine treatments (0.5 - 2 mM) clearly decreased the cell growth of Hep G2 and SK-Hep-1 cells to 72.5 ± 3.4% and 71.3 ± 4.6% of control, respectively. In parallel, adenine also induced sub-G1 and S phase accumulation in both HCC cells. However, adenine did not affect the cell growth and cell cycle distribution of Hep3B cell. Western blot analysis showed that adenine reduced expression of cyclin A/D1 and cyclin-dependent kinase (CDK)2 and upregulated p53, p21, Bax, PUMA, and NOXA in HepG2 cell. Moreover, adenine induced AMPK activation that was involved in the p53-associated apoptotic cascade in HepG2 cells. Inhibition of AMPK activation or knockdown of AMPK restored the decreased cell growth of HepG2 and SK-Hep-1 cells in response to adenine. Conclusions: These findings reveal that adenine reduces the cell growth of HepG2 and SK-Hep-1 but not Hep3B cells, attributing to the AMPK/p53-mediated S phase arrest and apoptosis. It suggests that adenine has anticancer potential against p53-wild type HCC cells and may be beneficial as an adjuvant for HCC treatment.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e56fdd3511f1363046bc8e0f89ad9d6eTest
http://europepmc.org/articles/PMC7085215Test

المؤلفون: You-Cian Lin, Shao-Hsuan Kao, Chia-Hung Hung, Han-Min Chen, Chau-Jong Wang, Jiun-Tsai Lin, Chien-Wei Huang

المصدر: Pharmaceuticals, Vol 14, Iss 860, p 860 (2021)
Pharmaceuticals
Volume 14
Issue 9

مصطلحات موضوعية: colorectal cancer cell, Cell, Integrin, Pharmaceutical Science, Article, Focal adhesion, Pharmacy and materia medica, Drug Discovery, medicine, Protein kinase A, adenine, PI3K/AKT/mTOR pathway, Paxillin, biology, AMP-activated protein kinase, Chemistry, focal adhesion kinase, AMPK, invasion, RS1-441, medicine.anatomical_structure, Cancer research, biology.protein, Molecular Medicine, Medicine, Proto-oncogene tyrosine-protein kinase Src

الوصف: Tumor metastasis is a major cause of death of patients with colorectal cancer (CRC). Our previous findings show that adenine has antiproliferation activity against tumor cells. However, whether adenine reduces the invasiveness of DLD-1 and SW480 CRC cells has not been thoroughly explored. In this study, we aimed to explore the effects of adenine on the invasion potential of DLD-1 cells. Our findings showed that adenine at concentrations of ≤200 μM did not influence the cell viability of DLD-1 and SW480 CRC cells. By contrast, adenine reduced the migratory potential of the CRC cells. Moreover, it decreased the invasion capacity of the CRC cells in a dose-dependent manner. We further observed that adenine downregulated the protein levels of tissue plasminogen activator, matrix metalloproteinase-9, Snail, TWIST, and vimentin, but upregulated the tissue inhibitor of metalloproteinase-1 expression in DLD-1 cells. Adenine decreased the integrin αV level and reduced the activation of integrin-associated signaling components, including focal adhesion kinase (FAK), paxillin, and Src in DLD-1 cells. Further observations showed that adenine induced AMP-activated protein kinase (AMPK) activation and inhibited mTOR phosphorylation in DLD-1 cells. The knockdown of AMPK restored the reduced integrin αV level and FAK/paxillin/Src signaling inhibited by adenine in DLD-1 cells. Collectively, these findings reveal that adenine reduces the invasion potential of DLD-1 cells through the AMPK/integrin/FAK axis, suggesting that adenine may have anti-metastatic potential in CRC cells.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::24dda4101ce91c5dad9bf50c50b6921dTest
https://www.mdpi.com/1424-8247/14/9/860Test

المؤلفون: Jyh-Gang Leu, Yen-Lin Chen, Ming-Hsing Chiang, Chao-Yi Chen, Yao-Jen Liang, Jiun-Tsai Lin, Han-Min Chen

المصدر: European journal of pharmacology. 818

مصطلحات موضوعية: 0301 basic medicine, Male, medicine.medical_specialty, Time Factors, Angiogenesis, Peroxisome proliferator-activated receptor, Neovascularization, Physiologic, AMP-Activated Protein Kinases, Cell Line, Diabetes Mellitus, Experimental, 03 medical and health sciences, Mice, 0302 clinical medicine, AMP-activated protein kinase, Internal medicine, Diabetes mellitus, Medicine, Animals, Humans, PPAR delta, Protein kinase A, Pharmacology, chemistry.chemical_classification, Wound Healing, biology, business.industry, Adenine, AMPK, medicine.disease, Enzyme Activation, 030104 developmental biology, Endocrinology, chemistry, 030220 oncology & carcinogenesis, biology.protein, Peroxisome proliferator-activated receptor delta, business, Wound healing, Signal Transduction

الوصف: Wound healing is one of the major complications of diabetes, and problems with wound healing in diabetics often lead to amputation and even death. AMP-activated protein kinase (AMPK) is a protein involved in intracellular metabolism. Activated AMPK can reduce visceral fat and cholesterol synthesis and even inhibit hepatic gluconeogenesis. Activation of AMPK has been widely used in the treatment of type II diabetes. We applied an AMPK activator (Adenine) to human fibroblasts and to the wounds of streptozotocin-induced diabetic mice. We applied Adenine ointment to the wounds on 7 consecutive days and observed the healing status as well as activation of AMPK and angiogenic factors. Based on the appearance of the wounds, the results showed that after 7 days of treatment the wound area was smaller in the Adenine-treated group relative to the control group. The results for tissue protein expression showed that, compared to the control group, angiogenic related protein, PPARδ were increased and receptor for advanced glycation endproducts (RAGE) was decreased in the Adenine-treated group. Our studies indicate that Adenine has the potential to become a useful drug in the treatment of diabetic wound healing.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b5a990c722a0dbab393c5ff765c26724Test
https://pubmed.ncbi.nlm.nih.gov/29162431Test

المؤلفون: Chun-Hsiang Lin, Jiun-Tsai Lin, Yi-Fang Cheng, Bee-Piao Huang, Shao-Hsuan Kao, Han-Min Chen

المصدر: DNA and Cell Biology. 34:133-141

مصطلحات موضوعية: Lipopolysaccharides, Adenosine monophosphate, Cell Survival, Immunoblotting, Down-Regulation, Gene Expression, AMP-Activated Protein Kinases, p38 Mitogen-Activated Protein Kinases, Cell Line, Nitric oxide, Proinflammatory cytokine, Mice, chemistry.chemical_compound, Genetics, Animals, Phosphorylation, Protein kinase A, Molecular Biology, biology, Plant Extracts, Reverse Transcriptase Polymerase Chain Reaction, Macrophages, NF-kappa B, AMPK, Interleukin, Cell Biology, General Medicine, Cell biology, Enzyme Activation, Plant Leaves, Nitric oxide synthase, chemistry, Immunology, biology.protein, Cytokines, lipids (amino acids, peptides, and proteins), Tumor necrosis factor alpha, Inflammation Mediators, Phosphatidylinositol 3-Kinase, Sasa, Signal Transduction

الوصف: Adenosine monophosphate (AMP)-activated protein kinase (AMPK) plays a central role in energy homeostasis and regulation of inflammatory responses. The present study is aimed to investigate the anti-inflammatory effects of ENERGI-F704, a nucleobase analogue isolated from bamboo leaves, on expression of proinflammatory mediators in murine macrophage RAW264.7 in response to lipopolysaccharide (LPS). ENERGI-F704 enhanced phosphorylation of AMPK(T172) but insignificantly affected the viability of RAW264.7 cells. Further investigation showed that ENERGI-F704 decreased mRNA expression of interleukin (IL)-6, IL-8, tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX2), and inducible nitric oxide synthase (iNOS) induced by LPS, as well as suppressed the production of prostaglandin E2 (PGE₂) and nitric oxide (NO). Additionally, the inhibitory effects of ENERGI-F704 on the LPS-induced proinflammatory mediators were diminished by pretreatment of AMPK inhibitor Compound C. ENERGI-F704 also inhibited LPS-triggered activation of nuclear factor kappa B (NF-κB), phosphatidylinositol 3-kinase (PI3K), and p38 mitogen-activated protein kinase (p38), whereas extracellular signal-regulated kinase (Erk)1/2 and c-Jun N-terminal kinase (JNK) were insignificantly influenced. Our findings indicate that ENERGI-F704 may exert anti-inflammatory activity on RAW264.7 cells in response to LPS through the activation of AMPK and suppression of PI3K/P38/NF-κB signaling and the consequent decreased expression of proinflammatory mediators, suggesting that ENERGI-F704 is beneficial to the amelioration of inflammatory disorders.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b4180501fb10c9d1a64b78e70a90a3cfTest
https://doi.org/10.1089/dna.2014.2630Test

المؤلفون: Han-Min Chen, Chia-Nan Chen, Chi-Shun Tu, Yao-Jen Liang, Po-Jung Chen, Shan-Wen Tan, Chao-Yi Chen, Yi-Huei Huang

المصدر: RSC Adv.. 4:4656-4662

مصطلحات موضوعية: biology, General Chemical Engineering, General Chemistry, Epigallocatechin gallate, Pharmacology, Bioavailability, chemistry.chemical_compound, chemistry, Colloidal gold, Drug delivery, Hyaluronic acid, biology.protein, Immunohistochemistry, Epidermal growth factor receptor, Wound healing

الوصف: Diabetic ulcers and unhealed bedsores have resulted in serious complications all around the world. Anti-oxidant epigallocatechin gallate (EGCG) has been proved beneficial in diabetic studies. However, the low bioavailability of EGCG is always a problem for human application. In this study, topical gas-injection of a EGCG and gold nanoparticle (AuNP) liquid mixture (AuE) using the GNT GoldMed™ Liquid Drug Delivery System significantly accelerated the wound healing on wild-type and streptozotocin-induced diabetic mouse skin. Immunoblotting of the diabetic wound tissue showed a significant increase of the vascular endothelial cell growth factor on day 7 and the Cu/Zn superoxide dismutase expression from day 3 to day 7. Furthermore, the epidermal growth factor receptor and collagen I & III protein expression both increased significantly in the wound area. After gas-injection of the AuE liquid, hyaluronic acid (HA) expression also significantly increased on day 7 as measured by immunohistochemistry analysis. In conclusion, gas-injection of AuE significantly increases the rate of wound healing both in wild-type and diabetic mice. This study may provide a new approach for improving the EGCG bioavailability on diabetic wounds.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::16109df31eb6c89a6625fa402f69f055Test
https://doi.org/10.1039/c3ra44359kTest

المؤلفون: Yao-Jen Liang, Han-Min Chen, Yi-Chuan Cheng, Chia-Hui Yen, Ming-Chang Chiang

المصدر: Mitochondrion. 14:7-17

مصطلحات موضوعية: Neurons, chemistry.chemical_classification, biology, Neurite, Sirtuin 1, Peroxisome proliferator-activated receptor, Cell Biology, TFAM, CREB, Molecular biology, Cell biology, PPAR gamma, Rosiglitazone, Mice, chemistry, Cell Line, Tumor, Coactivator, Neurites, biology.protein, Animals, Molecular Medicine, Thiazolidinediones, NRF1, Protein kinase A, Molecular Biology

الوصف: Several pieces of evidence indicate that peroxisome proliferator-activated receptor gamma (PPARγ) stimulation promotes neuronal differentiation. However, to date, the effects of a synthetic PPARγ agonist (Rosiglitazone, Rosi) on neurite outgrowth have not yet been well described. Here we have evaluated the effects of Rosi on neurite outgrowth and mitochondrial function in the mouse neuroblastoma Neuro 2a (N2A) cell line. Our results show that Rosi promotes neurite outgrowth of N2A cells and significantly increases the population of neurite-bearing cells, with apparent increase of intracellular calcium and the expression of calmodulin-dependent kinase I (CaMKI). Rosi also increases the intracellular cAMP and expression of both protein kinase A (PKA) and cAMP response element binding protein (CREB). Phosphorylation of CREB was also detected in the Rosi treated N2A cells. Moreover, Rosi significantly increases the transcription of AMP-activated kinase (AMPK) and Sirtuin 1 (SIRT1). Besides, the expression of PPAR coactivator 1α (PGC1α), as well as the mRNA level its downstream genes, including nuclear respiratory factors 1 and 2 (NRF1 and NRF2) and mitochondrial transcription factor A (Tfam) were induced by Rosi treatments. Furthermore, Rosi increases the level of ATP, D-loop, and mitochondrial mass in N2A cells. Collectively, these findings provide an array of evidence that PPARγ activation provides beneficial neuronal networks within neurite outgrowth.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::355baa9386a24483b99888821d92584cTest
https://doi.org/10.1016/j.mito.2013.12.003Test

المؤلفون: Ching-Yu Lin, Vinchi Wang, Han-Min Chen

المصدر: Clinical Biochemistry. 44:377-385

مصطلحات موضوعية: Male, Proteomics, medicine.medical_specialty, Parkinson's disease, Clinical Biochemistry, Enzyme-Linked Immunosorbent Assay, Mass Spectrometry, Central nervous system disease, Degenerative disease, Internal medicine, Blood plasma, medicine, Humans, Electrophoresis, Gel, Two-Dimensional, Two sample, Serum amyloid P component, Aged, Aged, 80 and over, biology, Chemistry, Parkinson Disease, General Medicine, Middle Aged, medicine.disease, Amyloid P Component, Serum Amyloid P-Component, Endocrinology, Case-Control Studies, biology.protein, Female, Biomarkers

الوصف: Objectives Parkinson's disease (PD) ranks the second among the neurodegenerative disorders. Proteins involved in Parkinson's disease (PD) have been investigated but none as the diagnostic markers in blood. Design and methods In this study, we applied a proteomic strategy, by utilizing two-dimensional electrophoresis and mass spectrometry, to analyze two sample pools of plasma from the healthy individuals and PD subjects. Results IgGκL and human serum amyloid P component (SAP) were found differentially expressed between these pools. SAP level increased by approximately 5-fold in PD samples, and the ELISA procedure revealed a significant (P

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0006f4db9c41b6d621f92c50611f5989Test
https://doi.org/10.1016/j.clinbiochem.2011.01.002Test

المؤلفون: Chi-Hong Chu, Jiann-Shing Wu, Yu-Jen Wu, Szu-Yu Wu, Ching-Yu Lin, Ren-Jeh Lee, Rong-Huay Juang, Han-Min Chen, Pei-Ru Huang, Li-Te Chin

المصدر: Journal of Immunological Methods. 361:89-97

مصطلحات موضوعية: medicine.drug_class, Immunology, Cell Culture Techniques, Monoclonal antibody, Cell Fusion, Mice, Plasma, Antibody Repertoire, Antigen, Blood plasma, medicine, Animals, Humans, Immunology and Allergy, Cell Proliferation, Mice, Inbred BALB C, Hybridomas, Cell fusion, biology, Cell growth, Antibodies, Monoclonal, Virology, Molecular biology, Cell culture, biology.protein, Antibody

الوصف: Prion diseases such as Bovine Spongiform Encephalopathy (BSE) and new variant Creutzfeldt-Jakob disease (nvCJD) have caused a major safety concern in cell cultures using fetal calf serum (FCS). In this study, we found that screened and tested human plasma (HP) obtained from blood centers may be an ideal alternate nutrient substitute to FCS for culturing hybridoma. In addition to the inherent safety, a ten-fold increase in the fusion efficiency has been observed if the HP was used as the nutrient supplement instead of FCS. Subsequently, a broader antibody repertoire may be recovered. The HP supplement was found to promote the growth of hybridoma cells but no impact on antibody secretion. Interestingly, this effect of enrichment was only observed for HP, but not plasma from other animals. Unidentified murine hybridoma cloning factors other than IL-6 may specifically reside in human blood.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fec919db23e893ca5f10ed5582e070c0Test
https://doi.org/10.1016/j.jim.2010.08.001Test

المؤلفون: Cheng-Di Chiu, Ai-Ling Hour, Chi-Hong Chu, Nai-Kuei Huang, Pei-Ru Huang, Kuang-Yu Hu, Li-Te Chin, Hao-Ai Shui, Han-Min Chen

المصدر: Journal of Proteome Research. 9:3091-3102

مصطلحات موضوعية: Proteomics, Drug Resistance, Kaplan-Meier Estimate, Biochemistry, Mice, chemistry.chemical_compound, Drug Stability, Cell Line, Tumor, Dihydrofolate reductase, Animals, Cluster Analysis, Humans, Blood Transfusion, Electrophoresis, Gel, Two-Dimensional, Cytotoxicity, Cell Proliferation, Retrospective Studies, Dose-Response Relationship, Drug, biology, Cell growth, Adenine, Systems Biology, Preservatives, Pharmaceutical, Translation (biology), Blood Proteins, General Chemistry, In vitro, Aminopterin, Up-Regulation, Cell biology, Biomarker, Methotrexate, chemistry, Blood Preservation, Antifolate, Immunology, biology.protein, Folic Acid Antagonists

الوصف: The inhibition of dihydrofolate reductase (DHFR) by antifolates is a common practice both in cell culture and in chemotherapy. Surprisingly, antifolate resistance was also observed in cultured murine myeloma cells (SP2/0) in the presence of human plasma (HP); thus, we used a proteomic approach to identify novel plasma biomarker(s) for this condition. In contrast to the in vitro antifolate response, metabolic enzymes and translation machinery proteins were found to be up-regulated in the presence of HP. The antifolate resistance inherent in HP may be explained by a simultaneous promotion of cell proliferation and the maintenance of DNA integrity. Furthermore, the factor(s) was found to be extrinsic, heat stable and very small in size. Adenine, a supplemented additive in erythrocyte preservation, was subsequently identified as the contributing factor and exogenous addition in cultures reversed the cytotoxicity induced by antifolates. Importantly, adenine-containing blood components, which may provide enhanced survival to otherwise sensitive antifolate-targeted cells, showed a dose-dependent adverse effect in transfusion recipients receiving antifolate (methotrexate) medications. These findings not only highlight a previously unnoticed role of adenine, but also emphasize a novel mechanistic link between transfusion and subsequently reduced survival in patients taking methotrexate.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::50d91a8207c85f8dcc78358d6ead95b3Test
https://doi.org/10.1021/pr100005uTest

المؤلفون: Jing-Yi Nong, Po-Ku Chen, Yi-Fang Cheng, Cheng-Yu Chao, Han-Min Chen, Jiun-Tsai Lin, Chun-Fang Huang, Guang-Huar Young

المصدر: Journal of proteomics. 120

مصطلحات موضوعية: biology, Proteome, Activator (genetics), Glucose uptake, Adenine, Biophysics, Glucose transporter, AMPK, AMP-Activated Protein Kinases, Biochemistry, Gene Expression Regulation, Enzymologic, Enzyme Activation, Insulin receptor, Mice, Glucose, Downregulation and upregulation, biology.protein, NIH 3T3 Cells, Animals, Glycolysis, Protein kinase A

الوصف: AMP-activated protein kinase (AMPK) is a metabolic master switch maintaining the energy homeostasis in cells and thought to modulate cellular response to stresses. Adenine as well as a pharmacological AMPK activator, 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR), induced the phosphorylation of AMPK and acetyl-CoA carboxylase in NIH/3T3 cells. Administration of adenine or AICAR increased the expression and translocation of glucose transporter 4, enhanced the cellular glucose uptake, and elevated the intracellular ATP level. To better understand the proteomic changes in response to exogenous adenine treatment, we performed two-dimensional difference gel electrophoresis (2DE-DIGE) and grouped protein spots with similar intensities prior to MS analysis. These process allowed us to exclude these constant expressed proteins, reduce the coverage from abundant signals and increase the identification of middle/lower expressed proteins. Bioinformatics analysis on the proteomic alterations suggested that both of adenine and AICAR could induce up-regulation of a panel of proteins associated with glucose metabolism. We also found that adenine upregulated expression of the glycolytic enzyme, hexokinase 2, indicating a link between adenine and AMPK-mediated glycolysis. Taken together, by demonstrating the adenine-mediated proteome changes in NIH/3T3 cells, our study provides useful information for the characteristics of adenine-induced AMPK activation and development of efficient AMPK activator. Biological significance AMPK is a fuel sensing enzyme that responds to a central role of energy homeostasis and contributes to the acceleration of insulin signaling. Recently, we have shown that exogenous adenine exerted anti-inflammatory effects through activation of AMPK, suggesting the treatment is a potent therapeutic strategy against hyperglycemia. Adenine had similar effects with 5-amino-4-imidazole-carboxamide riboside (AICAR, an AMPK activator) in modulating glucose uptake via AMPK-mediated signaling. In this study, we performed a 2DE-DIGE/MS-based approach to investigate the mechanism of exogenous adenine in NIH/3T3 cells. Our results provide evidence of a novel role for adenine in AMPK-mediated signaling and glucose metabolism and suggest potential therapeutic perspectives in insulin resistance and metabolic dysfunctions.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d152508f18fbf3b0be0fa5a2bec291b8Test
https://pubmed.ncbi.nlm.nih.gov/25797921Test