RESUMO
Protein kinase CK2 is active in cancer cells. Previously, we reported that increased CK2 activity could induce epithelial mesenchymal transition of cancer cells. CK2 also induced epithelial mesenchymal transition in colon cancer cell lines such as HT29 and SW620, and the transitioned cells (CK2α cells) became more proliferative than the controls. We assumed that CK2 could affect cancer cell growth by modulating their energy metabolism. Here, we examined the molecular effects of CK2 on the glucose metabolism of cancer cells. We found that CK2α cells consumed more glucose and produced more lactate than control cells did. An XF glycolysis stress test showed that aerobic glycolysis was augmented up to the cancer cell's maximal glycolytic capacity in CK2α cells. Molecular analysis revealed that pyruvate kinase M1 was downregulated and pyruvate kinase M2 was nuclear localized in CK2α cells. Consequently, the expression and activity of lactate dehydrogenase A (LDHA) were upregulated. Treatment with FX11-a specific LDHA inhibitor-or clustered regularly interspaced short palindromic repeats (CRISPR)-mediated knockout of LDHA inhibited the CK2-driven proliferation of cancer cells. We conclude that CK2 augments the Warburg effect, resulting in increased proliferation of cancer cells.
Assuntos
Neoplasias do Colo/enzimologia , Neoplasias do Colo/patologia , Glicólise , Piruvato Quinase/genética , Piruvato Quinase/metabolismo , Caseína Quinase II/genética , Caseína Quinase II/metabolismo , Proliferação de Células , Transição Epitelial-Mesenquimal , Regulação Neoplásica da Expressão Gênica , Técnicas de Inativação de Genes , Glucose/metabolismo , Células HEK293 , Células HT29 , Humanos , Isoenzimas/metabolismo , Lactato Desidrogenase 5/antagonistas & inibidores , Lactato Desidrogenase 5/genética , Lactato Desidrogenase 5/metabolismo , Ácido Láctico/metabolismo , Naftalenos/farmacologia , Ativação TranscricionalRESUMO
ω-Hydroxyundec-9-enoic acid (ω-HUA), a plant secondary metabolite, exhibits anti-fungal activity. However, its effect on breast cancer cells is unknown. Here, we investigated the anti- breast cancer activity of ω-HUA and its underlying mechanism. Treatment of human breast cancer cell lines, MDA-MB-231 and MDA-MB-435, with ω-HUA induced apoptotic cell death with increased cleaved caspase-3 and poly (ADP-ribose) polymerase (PARP) levels, and p38 and JNK phosphorylation. Inhibition of these mitogen-activated protein kinase (MAPK) pathways using specific inhibitors or siRNA, for p38 and JNK, respectively, blocked the ω-HUA-induced apoptosis in a dose-dependent manner. Moreover, pretreatment of the cells with antioxidant N-acetyl cysteine (NAC) inhibited ω-HUA-induced increased reactive oxygen species (ROS) levels, cleaved caspase-3 and cleaved PARP, and phosphorylated JNK, phosphorylated p38, and increased cell viability and colony-forming ability. MDA-MB-231 xenograft model showed that the ω-HUA-treated group exhibited greater tumor regression and significantly reduced tumor weight compared to that exhibited by the vehicle-administered group. Collectively, ω-HUA-induced intracellular ROS generation induced breast cancer cell apoptosis through JNK and p38 signaling pathway activation, resulting in tumor regression. The results suggested that ω-HUA is an effective supplement for inhibiting human breast cancer growth.
Assuntos
Antineoplásicos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , MAP Quinase Quinase 4/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Ácidos Undecilênicos/administração & dosagem , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Antineoplásicos/farmacologia , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Camundongos , Fosforilação/efeitos dos fármacos , Ácidos Undecilênicos/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
OBJECTIVES: Nanoparticulation using fat and supercritical fluid (NUFSTM) is a drug delivery platform technology enabling efficient and effective formulation of poorly soluble drugs. We performed experiments to examine whether NUFS™ could improve poor bioavailability and reduce fed-fasted bioavailability variances of erlotinib (Ert). METHODS: NUFS-Ert was prepared using NUFS™ technology; its physical properties were characterized, and drug release was measured. Furthermore, in vitro and in vivo efficacy tests and pharmacokinetic analysis were performed. RESULTS: NUFS-Ert nanoparticles had an average size of 250 nm and were stable for 2 months at 40 °C, 4 °C, and room temperature. The dissolution rate of NUFS-Ert increased in bio-relevant dissolution media. NUFS-Ert was more potent in inhibiting EGF signaling and in suppressing the proliferation of A549, a human non-small cell lung cancer cell line. Furthermore, A549 xenografts in BALB/c nude mice treated with NUFS-Ert regressed more efficiently than those in the mice treated with vehicle or Tarceva®. In addition, experimental lung metastasis was more efficiently inhibited by NUFS-Ert than by Tarceva®. The relative bioavailability of NUFS-Ert compared with that of Tarceva® was 550% and the ratio of the area under the concentration-time curve (AUC) of fed state to the AUC of fasted state was 1.8 for NUFS-Ert and 5.8 for Tarceva®. CONCLUSIONS: NUFS-Ert could improve poor bioavailability and reduce fed-fasted bioavailability variances of Ert. NUFS-Ert was more efficacious than Tarceva®.
Assuntos
Antineoplásicos/farmacocinética , Disponibilidade Biológica , Cloridrato de Erlotinib/farmacocinética , Excipientes/química , Neoplasias Pulmonares/tratamento farmacológico , Nanopartículas/química , Animais , Antineoplásicos/química , Química Farmacêutica , Cloridrato de Erlotinib/química , Cloridrato de Erlotinib/farmacologia , Humanos , Camundongos Nus , SolubilidadeRESUMO
ω-Hydroxyundec-9-enoic acid (ω-HUA), a hydroxyl unsaturated fatty acid derivative, is involved in the antifungal activity of wild rice (Oryza officinalis). Here, we investigated the anti-cancer activity of ω-HUA on a non-small cell lung cancer (NSCLC) cell line. ω-HUA increased apoptosis and induced cleavages of caspase-6, caspase-9, and poly (ADP-ribose) polymerase (PARP). ω-HUA treatment significantly induced endoplasmic reticulum (ER) stress response. Suppression of CHOP expression and inhibiting ER stress by 4-phenylbutyrate (4-PBA) significantly attenuated the ω-HUA treatment-induced activation of caspase-6, caspase-9, and PARP, and subsequent apoptotic cell death, indicating a role for ER stress in ω-HUA-induced apoptosis. In addition, cells subjected to ω-HUA exhibited significantly increased quantity of reactive oxygen species (ROS), and the ROS scavenger N-acetyl-L-cysteine (NAC) inhibited ω-HUA-induced apoptotic cell death and ER stress signals, indicating a role for ROS in ER stress-mediated apoptosis in ω-HUA-treated cells. Taken together, these results suggest that sequential ROS generation and ER stress activation are critical in ω-HUA treatment-induced apoptosis and that ω-HUA represents a promising candidate for NSCLC treatment.
Assuntos
Antineoplásicos/farmacologia , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Ácidos Undecilênicos/farmacologia , Acetilcisteína/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Caspase 6/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Técnicas de Silenciamento de Genes , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Poli(ADP-Ribose) Polimerases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição CHOP/antagonistas & inibidores , Fator de Transcrição CHOP/genética , Fator de Transcrição CHOP/metabolismoRESUMO
Cellular responses of Saccharomyces cerevisiae to high temperatures of up to 42 °C during ethanol fermentation at a high glucose concentration (i.e., 100 g/L) were investigated. Increased temperature correlated with stimulated glucose uptake to produce not only the thermal protectant glycerol but also ethanol and acetic acid. Carbon flux into the tricarboxylic acid (TCA) cycle correlated positively with cultivation temperature. These results indicate that the increased demand for energy (in the form of ATP), most likely caused by multiple stressors, including heat, acetic acid, and ethanol, was matched by both the fermentation and respiration pathways. Notably, acetic acid production was substantially stimulated compared to that of other metabolites during growth at increased temperature. The acetic acid produced in addition to ethanol seemed to subsequently result in adverse effects, leading to increased production of reactive oxygen species. This, in turn, appeared to cause the specific growth rate, and glucose uptake rate reduced leading to a decrease of the specific ethanol production rate far before glucose depletion. These results suggest that adverse effects from heat, acetic acid, ethanol, and oxidative stressors are synergistic, resulting in a decrease of the specific growth rate and ethanol production rate and, hence, are major determinants of cell stability and ethanol fermentation performance of S. cerevisiae at high temperatures. The results are discussed in the context of possible applications.
Assuntos
Ácido Acético/metabolismo , Etanol/metabolismo , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efeitos da radiação , Trifosfato de Adenosina/metabolismo , Carbono/metabolismo , Metabolismo Energético , Fermentação , Glucose/metabolismo , Análise do Fluxo Metabólico , Saccharomyces cerevisiae/crescimento & desenvolvimento , TemperaturaRESUMO
Chronic oral inflammation and biofilm-mediated infections drive diseases such as dental caries and periodontitis. This study investigated the anti-inflammatory and antibacterial potential of an ethanol extract from Astilbe chinensis inflorescence (GA-13-6) as a prominent candidate for natural complex substances (NCS) with therapeutic potential. In LPS-stimulated RAW 264.7 macrophages, GA-13-6 significantly suppressed proinflammatory mediators, including interleukin-6 (IL-6), tumor necrosis factor (TNF), and nitric oxide (NO), surpassing purified astilbin, a known bioactive compound found in A. chinensis. Furthermore, GA-13-6 downregulated the expression of cyclooxygenase-2 (COX2) and inducible nitric oxide synthase (iNOS), indicating an inhibitory effect on the inflammatory cascade. Remarkably, GA-13-6 exhibited selective antibacterial activity against Streptococcus mutans, Streptococcus sanguinis, and Porphyromonas gingivalis, key players in dental caries and periodontitis, respectively. These findings suggest that complex GA-13-6 holds the potential for the treatment or prevention of periodontal and dental diseases, as well as various other inflammation-related conditions, while averting the induction of antibiotic resistance.
Assuntos
Macrófagos , Extratos Vegetais , Animais , Camundongos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Células RAW 264.7 , Antibacterianos/farmacologia , Inflamação/tratamento farmacológico , Etanol/química , Óxido Nítrico Sintase Tipo II/metabolismo , Anti-Inflamatórios/farmacologia , Inflorescência/química , Ciclo-Oxigenase 2/metabolismo , Ciclo-Oxigenase 2/genética , Óxido Nítrico/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: We sought to quantify diffuse parenchymal lung disease (DPLD) extent using quantitative computed tomography (CT) analysis and to investigate its association with radiation pneumonitis (RP) development in non-small cell lung cancer (NSCLC) patients receiving definitive concurrent chemoradiation therapy (CCRT). METHODS: A total of 82 NSCLC patients undergoing definitive CCRT were included in this prospective cohort study. Pretreatment CT scans were analyzed using quantitative CT analysis software. Low-attenuation area (LAA) features based on lung density and texture features reflecting interstitial lung disease (ILD) were extracted from the whole lung. Clinical and dosimetric factors were also evaluated. RP development was assessed using the Common Terminology Criteria for Adverse Events version 5.0. Univariable and multivariable logistic regression analyses were performed to identify independent risk factors for grade ≥3 (≥GR3) RP. RESULTS: RP was identified in 68 patients (73.9%), with nine patients (10.9%) experiencing ≥GR3 RP. Univariable logistic regression analysis identified excess kurtosis and high-attenuation area (HAA)_volume (cc) as significantly associated with ≥GR3 RP. Multivariable logistic regression analysis showed that the combined use of imaging features and clinical factors (forced expiratory volume in 1 second [FEV1], forced vital capacity [FVC], and CHEMO regimen) demonstrated the best performance (area under the receiver operating characteristic curve = 0.924) in predicting ≥GR3 RP. CONCLUSION: Quantified imaging features of DPLD obtained from pretreatment CT scans would predict the occurrence of RP in NSCLC patients undergoing definitive CCRT. Combining imaging features with clinical factors could improve the accuracy of the predictive model for severe RP.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Doenças Pulmonares Intersticiais , Neoplasias Pulmonares , Pneumonite por Radiação , Humanos , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/radioterapia , Pneumonite por Radiação/etiologia , Pneumonite por Radiação/epidemiologia , Neoplasias Pulmonares/tratamento farmacológico , Estudos Prospectivos , Doenças Pulmonares Intersticiais/diagnóstico por imagem , Doenças Pulmonares Intersticiais/complicações , Estudos RetrospectivosRESUMO
Saccharomyces cerevisiae is an excellent ethanol producer, but is rather sensitive to high concentration of ethanol. Here, influences of ethanol on cellular membrane integrity and carbon metabolism of S. cerevisiae were investigated to rationalize mechanism involved in ethanol toxicity. Addition of 5% (v/v) ethanol did neither significantly change the permeability of the cytoplasmic membrane of the reference strain S. cerevisiae BY4741 nor of the ethanol-tolerant strain iETS3. However, the addition of ethanol resulted in a marked decrease in the mitochondrial membrane potential and in increased concentrations of intracellular reactive oxygen species (ROS). The carbon flux was redistributed under these conditions from mainly ethanol production to the TCA cycle. This redistribution was possibly a result of increased energy demand for cell maintenance that increased from about zero to 20-40 mmol ATP (g(CDW) h)(-1) . This increase in maintenance energy might be explained by the ethanol-induced reduction of the proton motive force and the required removal of ROS. Thus, the stability of the mitochondrial membrane and subsequently the capacity to keep ROS levels low could be important factors to improve tolerance of S. cerevisiae against ethanol.
Assuntos
Carbono/metabolismo , Etanol/toxicidade , Membranas Mitocondriais/efeitos dos fármacos , Saccharomyces cerevisiae/efeitos dos fármacos , Trifosfato de Adenosina/metabolismo , Permeabilidade da Membrana Celular/efeitos dos fármacos , Ciclo do Ácido Cítrico , Metabolismo Energético/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Membranas Mitocondriais/fisiologia , Saccharomyces cerevisiae/fisiologiaRESUMO
The effects of structural modification of cell wall on the biotransformation capability by recombinant Corynebacterium glutamicum cells, expressing the chnB gene encoding cyclohexanone monooxygenase of Acinetobacter calcoaceticus NCIMB 9871, were investigated. Baeyer-Villiger oxygenation of 2-(2'-acetoxyethyl) cyclohexanone (MW 170 Da) into R-7-(2'-acetoxyethyl)-2-oxepanone was used as a model reaction. The whole-cell biotransformation followed Michaelis-Menten kinetics. The V (max) and K (S) values were estimated as 96.8 U g(-1) of dry cells and 0.98 mM, respectively. The V (max) was comparable with that of cyclohexanone oxygenation, whereas the K (S) was almost eightfold higher. The K (S) value of 2-(2'-acetoxyethyl) cyclohexanone oxygenation was reduced by ca. 30% via altering the cell envelop structure of C. glutamicum with ethambutol, which inhibits arabinosyl transferases involved in the biosynthesis of cell wall arabinogalactan and mycolate layers. The higher whole-cell biotransformation rate was also observed in the oxygenation of ethyl 2-cyclohexanone acetate upon ethambutol treatment of the recombinant C. glutamicum. Therefore, it was assumed that the biotransformation efficiency of C. glutamicum-based biocatalysts, with respect to medium- to large-sized lipophilic organic substrates (MW > ca. 170), can be enhanced by engineering their cell wall outer layers, which are known to function as a formidable barrier to lipophilic molecules.
Assuntos
Acinetobacter/enzimologia , Membrana Celular/enzimologia , Corynebacterium glutamicum/enzimologia , Cicloexanonas/metabolismo , Etambutol/metabolismo , Melhoramento Genético/métodos , Oxigenases/metabolismo , Acinetobacter/genética , Biotransformação/fisiologia , Membrana Celular/genética , Corynebacterium glutamicum/genética , Oxigenases/genéticaRESUMO
Lactobacillus reuteri AN417 is a newly characterized probiotic strain. The activity of AN417 against oral pathogenic bacteria is unknown. We investigated the antibacterial activity of cell-free L. reuteri AN417 culture supernatant (LRS) against three oral pathogens: Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus mutans. P. gingivalis and F. nucleatum have been implicated in periodontal disease, whereas S. mutans causes dental caries. Exposing these oral pathogenic bacteria to LRS significantly reduced their growth rates, intracellular ATP levels, cell viability, and time-to-kill. The minimal inhibitory volume of LRS was 10% (v/v) against P. gingivalis, 20% (v/v) for F. nucleatum, and 30% (v/v) for S. mutans. LRS significantly reduced the integrity of biofilms and significantly suppressed the expression of various genes involved in P. gingivalis biofilm formation. The L. reuteri AN417 genome lacked genes encoding reuterin, reuteran, and reutericyclin, which are major antibacterial compounds produced in L. reuteri strains. LRS treated with lipase and α-amylase displayed decreased antibacterial activity against oral pathogens. These data suggest that the antibacterial substances in LRS are carbohydrates and/or fatty acid metabolites. Our results demonstrate that LRS has antimicrobial activity against dental pathogenic bacteria, highlighting its potential utility for the prevention and treatment of P. gingivalis periodontal disease.
Assuntos
Antibacterianos/farmacologia , Fusobacterium nucleatum/efeitos dos fármacos , Limosilactobacillus reuteri/metabolismo , Porphyromonas gingivalis/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/metabolismo , Biofilmes/efeitos dos fármacos , Meios de Cultura/química , Meios de Cultura/metabolismo , Ácidos Graxos/metabolismo , Fezes/microbiologia , Fusobacterium nucleatum/fisiologia , Humanos , Limosilactobacillus reuteri/classificação , Limosilactobacillus reuteri/isolamento & purificação , Lipase/metabolismo , Testes de Sensibilidade Microbiana , Doenças Periodontais/microbiologia , Doenças Periodontais/patologia , Filogenia , Porphyromonas gingivalis/fisiologia , Streptococcus mutans/fisiologia , Suínos , alfa-Amilases/metabolismoRESUMO
The nature of the molecules underlying the radioresistance phenotype of laryngeal cancer cells remains to be established. We initially generated radioresistant laryngeal cancer cell lines from human HEp-2 cells with fractionated radiation. These RR-HEp-2 cells and isolated clones displayed more radioresistant and anti-apoptotic phenotypes than parental HEp-2 cells after radiation. Characteristics of RR-Hep-2 cell lines were confirmed by upregulation of radioresistance-related genes, such as epidermal growth factor receptor, Hsp90, and Bcl-xl. Subsequently, we examined proteome changes between HEp-2 and RR-HEp-2 cells and identified 16 proteins showing significantly altered expression levels. Interestingly, protein expression of chloride intracellular channel 1 (CLIC1) was markedly suppressed in RR-HEp-2 cells, compared with non-irradiated control cells. Suppression of CLIC1 with an indanyloxyacetic acid-94 or small interfering RNA led to radioresistance in HEp-2 cells by suppressing the radiation-induced cellular ROS level. However, ectopic overexpression of CLIC1 induced radiosensitivity in RR-HEp-2 cells via induction of ROS level after radiation, suggesting that the protein acts as a positive regulator of ROS production. Our results collectively indicate that suppression of CLIC1 contributes to acquisition of the radioresistance phenotype of laryngeal cancer cells via inhibition of ROS production, implying that this protein is an important candidate molecule for radiotherapy in radioresistant laryngeal cancer cells.
Assuntos
Canais de Cloreto/metabolismo , Proteômica/métodos , Tolerância a Radiação , Espécies Reativas de Oxigênio/metabolismo , Morte Celular/efeitos da radiação , Linhagem Celular Tumoral , Canais de Cloreto/antagonistas & inibidores , Humanos , Proteínas de Neoplasias/metabolismo , Tolerância a Radiação/efeitos da radiação , Radiação Ionizante , Reprodutibilidade dos TestesRESUMO
Chemotherapy regimens for non?small cell lung cancer (NSCLC) have various adverse effects on the human body. For this reason, probiotics have received attention regarding their potential value as a safe and natural complementary strategy for cancer prevention. This study analyzed the anticancer effects of aqueous extracts of probiotic bacteria Bifidobacterium bifidum (BB), Bifidobacterium longum (BL), Bifidobacterium lactis (BLA), Bifidobacterium infantis 1 (BI1), and Bifidobacterium infantis 2 (BI2) on NSCLC cell lines. When the aqueous extracts of probiotic Bifidobacterium species were applied to the NSCLC cell lines A549, H1299, and HCC827, cell death increased considerably; in particular, the aqueous extracts from BB and BLA markedly reduced cell proliferation. p38 phosphorylation induced by BB aqueous extract increased the expression of cleaved caspase 3 and cleaved poly (ADP-ribose) polymerase (PARP), consequently inducing the apoptosis of A549 and H1299 cells. When the p38 inhibitor SB203580 was applied, phosphorylation of p38 decreased, and the expression of cleaved caspase 3 and cleaved PARP was also inhibited, resulting in a reduction of cell death. In addition, BB aqueous extracts reduced the secretion of MMP-9, leading to inhibition of cancer cell invasion. By contrast, after transfection of short hairpin RNA shMMP-9 (for a knockdown of MMP-9) int°Cancer cells, BB aqueous extracts treatment failed to suppress the cancer cell invasiveness. According to our results about their anticancer effects on NSCLC, probiotics consisting of Bifidobacterium species may be useful as adjunctive anticancer treatment in the future.
Assuntos
Apoptose/efeitos dos fármacos , Bifidobacterium , Produtos Biológicos/farmacologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Probióticos/farmacologiaRESUMO
This study was performed to evaluate the anticancer effect of ω-hydroxyundec-9-enoic acid (ω-HUA), a microbial bio-catalyst product in breast cancer cells, through AMP-activated protein kinase (AMPK) regulation. ω-HUA mediated apoptosis was induced in breast cancer cells by AMPK activation, loss of mitochondrial membrane potential, and reactive oxygen species (ROS) generation. ω-HUA treatment of breast cancer cells increased the AMPK phosphorylation levels, cleaved caspase-3, and poly (ADP-ribose) polymerase (PARP) proteins. In addition, anti-apoptotic members, such as Bcl-2, were downregulated, while Bax, a pro-apoptotic member, was upregulated. ω-HUA decreased the mitochondrial membrane potential while increasing the expression of cytochrome c (cyt c). Treating the cells with compound C, an AMPK inhibitor, reversed the phenomena, leading to an increase in cell viability and a decrease in apoptosis induction. Treating the cells with an ROS scavenger, N-acetyl cysteine (NAC), led to AMPK inactivation and apoptosis inhibition, allowing the recovery of cell health. In conclusion, ω-HUA sequentially caused the production of mitochondrial ROS and the consequent AMPK activation, thereby inducing apoptosis in breast cancer cells. Thus, ω-HUA may prove useful as an anticancer agent that targets AMPK in breast cancer cells.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias da Mama/tratamento farmacológico , Espécies Reativas de Oxigênio/metabolismo , Ácidos Undecilênicos/farmacologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Fosforilação/efeitos dos fármacos , Espécies Reativas de Oxigênio/análise , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
The purpose of this study is to investigate the effect of Agrimonia pilosa Ledeb. extract (APLE) on lipopolysaccharide- (LPS-) induced cell damage in hepatocytes with a focus on antioxidant and anti-inflammatory activities. Total antioxidant and anti-inflammatory activities of APLE itself were analyzed and phytochemical analysis was performed. Moreover, inhibitory effects of APLE on LPS-induced oxidative stress and inflammation were assessed in human HepG2 hepatocytes. APLE was found to exert α,α-diphenyl-ß-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and nitrite scavenging activities and reducing power in a dose-dependent manner. The total phenolic and flavonoid contents of APLE were 44.30 ± 1.61 mg GAE/g and 29.65 ± 1.81 mg QE/g, respectively. HPLC analysis revealed that gallic acid is the major phenolic compound in APLE, followed by rutin, genistein, taxifolin, quercetin, luteolin, and apigenin, in descending order. Treatment of 100 and 200 µg/mL APLE significantly reduced LPS-stimulated intracellular reactive oxygen species production to the basal level without any cytotoxicity. Oppositely, APLE reversed LPS-suppressed expression of glutathione peroxidase gene and protein. Consistent with this result, APLE suppressed LPS-triggered expression of proinflammatory cytokine genes in a dose-dependent manner. These results reinforce the fact that the antioxidant and anti-inflammatory activity of APLE helps protect hepatocytes from LPS. Thus, APLE may be utilized as a bioactive ingredient in functional foods.
RESUMO
We investigated the intracellular metabolic fluxes of protein kinase CK2-activating (Cα OE) cells and role of lactate dehydrogenase A (LDHA) as a contributor of tumorigenesis after reprogrammed glucose metabolism. Facilitated aerobic glycolysis was confirmed via isotope tracer analysis, in which 13C6-Glc or 13C5-Gln was added to the media, following which metabolites converted from Cα OE cells were identified. We found a greater decrease in cell survival, colony-forming ability, migration, and Cα OE cell invasion under glucose (Glc)-depletion conditions than under glutamine (Gln)-depletion conditions. Cancer cell migration and invasion increased due to LDHA elevation of the altered metabolic axis driven by activated CK2. FX11 treatment and LDHA knockdown suppressed migration and invasion through ROS generation, but this was partially reversed by the antioxidant N-acetylcysteine (NAC). Moreover, LDHA inhibition decreased tumor growth in a mouse xenograft model transplanted with Cα OE cells. Finally, we concluded that LDHA is an excellent metabolic target for tumor therapy, based on CK2α derived aerobic glycolysis.
Assuntos
Caseína Quinase II/metabolismo , Glucose/metabolismo , Lactato Desidrogenases/metabolismo , Neoplasias/metabolismo , Aerobiose , Animais , Caseína Quinase II/genética , Ciclo Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Ciclo do Ácido Cítrico , Glicólise , Humanos , Camundongos , Neoplasias/patologia , Ácido Pirúvico/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Diabetes and obesity represent the major health problems and the most age-related metabolic diseases. Protein-tyrosine phosphatase 1B (PTP1B) has emerged as an important regulator of insulin signal transduction and is regarded as a pharmaceutical target for metabolic disorders. To find novel natural materials presenting therapeutic activities against diabetes and obesity, we screened various herb extracts using a chip screening allowing the determination of PTP1B inhibitory effects of the tested compounds using insulin receptor (IR) as the substrate. Cudrania tricuspidata leaves (CTe) had a strong inhibitory effect on PTP1B activity and substantially inhibited fat accumulation in 3T3-L1 cells. CTe was orally administrated to diet-induced obesity (DIO) mice once daily for 3 weeks after which changes in glucose, insulin metabolism, and fat accumulation were examined. Hepatic enzyme markers (aspartate aminotransferase, AST, and alanine aminotransferase, ALT) and total fat mass and triglyceride levels decreased in CTe-treated mice, whereas body weight and total cholesterol concentration slightly decreased. CTe increased the phosphorylation of IRS-1 and Akt in liver tissue. Furthermore, CTe treatment significantly lowered blood glucose levels and improved insulin secretion in DIO mice. Our results strongly suggest that CTe may represent a promising therapeutic substance against diabetes and obesity.
Assuntos
Diabetes Mellitus/tratamento farmacológico , Obesidade/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Proteína Tirosina Fosfatase não Receptora Tipo 1/biossíntese , Células 3T3-L1 , Animais , Diabetes Mellitus/genética , Diabetes Mellitus/patologia , Humanos , Insulina/sangue , Resistência à Insulina/genética , Fígado/efeitos dos fármacos , Fígado/patologia , Camundongos , Camundongos Obesos , Moraceae/química , Obesidade/genética , Obesidade/patologia , Extratos Vegetais/química , Transdução de Sinais/efeitos dos fármacosRESUMO
PURPOSE: The purpose of the study was to investigate fatigue, depression and sleep in young adult and middle-aged. METHOD: The convenient sample consisted of 415 subjects from 20 to 59 years old. Data were collected using a self-report questionnaire from July to October, 2001. The VAS-F and CES-D were utilized to measure the level of fatigue and depression. Sleep duration and sleep satisfaction were measured based on the subject's self-report. RESULT: The result of the study revealed that the level of fatigue and depression was higher among young adult than middle-aged. Considering age and gender, the level of fatigue and depression was higher among young adult women and middle-aged men. Depression and sleep satisfaction influenced on the fatigue. CONCLUSION: Health care providers need to concern about fatigue and depression in young adult women and middle-aged men. Especially, more concern and intervention programs are needed for young adult women and middle-aged men.
RESUMO
Small-sized particles are more suitable for targeted delivery and are therapeutically more effective than large-sized particles. In this study, we investigated the anticancer effects of ultrafine particles of Ulmus davidiana var. japonica (ufUJ) on human gastric cancer cell lines SNU-1, SNU-216, and SNU-484. ufUJ induced apoptosis by the proteolytic activation of caspase-9, caspase-6, and caspase-3 and cleavage of poly (ADP-ribose) polymerase. The expression levels of the endoplasmic reticulum stress-related protein BiP markedly increased after ufUJ treatment. BiP knockdown decreased ufUJ-induced cell death. ufUJ-induced apoptosis was inhibited by the caspase-3 inhibitor z-DEVD-fmk, caspase-6 inhibitor z-VEID-fmk, and caspase-9 inhibitor z-LEHD-fmk, and by siRNAs against caspases 3, 6, and 9. Gastric cancer cells did not show anchorage-independent growth in the presence of ufUJ. However, cells treated with caspase inhibitors showed an enhanced colony-forming ability. These findings may be helpful in the prevention of gastric cancer and in the development of functional foods.
Assuntos
Apoptose/fisiologia , Caspases/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Extratos Vegetais/farmacologia , Neoplasias Gástricas/metabolismo , Ulmus , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Relação Dose-Resposta a Droga , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Humanos , Técnicas de Cultura de Órgãos , Tamanho da Partícula , Extratos Vegetais/isolamento & purificaçãoRESUMO
The medicinal plant Ulmus davidiana var. japonica has significant potential as a cancer chemoprevention agent. Catechin-7-O-xyloside (C7Ox) was purified from ultrafine U. davidiana var. japonica ethanol extract. In the present study, we investigated the apoptotic effect of C7Ox in the non-small cell lung cancer (NSCLC) cell line H1299. C7Ox treatment induced cell death and decreased plasma membrane integrity, an event typical of apoptosis. C7Ox-induced apoptosis was associated with the proteolytic activation of caspase-6, cleavage of poly(ADP-ribose) polymerase (PARP) and loss of mitochondrial membrane potential. C7Ox also induced the endoplasmic reticulum (ER) stress-regulated pro-apoptotic transcription factor CHOP. The suppression of CHOP expression significantly decreased C7Ox-induced cell death, LDH leakage and caspase-6 activation. Antitumor effects, evaluated based on protracted tumor regression, were observed when nude-mice bearing H1299 xenografts were treated with C7Ox. C7Ox-induced tumor regression was accompanied by enhanced expression of CHOP mRNA. Our data suggest that C7Ox can trigger mitochondrial-mediated apoptosis, and that ER stress is critical for C7Ox-induced apoptosis in H1299 NSCLC cells.
Assuntos
Apoptose/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Caspase 6/biossíntese , Catequina/análogos & derivados , Catequina/farmacologia , Estresse do Retículo Endoplasmático , Neoplasias Pulmonares/enzimologia , Mitocôndrias/efeitos dos fármacos , Xilose/análogos & derivados , Animais , Linhagem Celular Tumoral , Humanos , L-Lactato Desidrogenase/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Poli(ADP-Ribose) Polimerases/metabolismo , Interferência de RNA , RNA Mensageiro/biossíntese , RNA Interferente Pequeno , Fator de Transcrição CHOP/biossíntese , Fator de Transcrição CHOP/genética , Xilose/farmacologiaRESUMO
PURPOSE: The purpose of this study was to explore the effects of a Taegyo program on parents-fetal attachment and parenthood in first pregnancy couples (mothers and spouses). METHODS: The research design was a nonequivalent control group pretest-posttest experiment. Study participants were 52 first pregnancy couples visiting two medium-scale obstetrics and gynecology clinics located in Gwangju. A total of 52 couples were assigned to the experimental group (25 couples) and the control group (27 couples). The experimental couples were provided with a Taegyo program for 4 weeks. Data were analyzed by chi square test, t-test, and ANCOVA using the SPSS program. RESULTS: Post-treatment maternal-fetal attachment, paternal-fetal attachment and motherhood significantly increased in the experimental group as compared to the control group, but post-treatment fatherhood, anxiety, blood pressure and pulse of participants in the experimental group showed no significant difference from those in the control group. CONCLUSION: From these results, it is suggested that the Taegyo program has beneficial effects in enhancing parent-fetal attachment and motherhood in first pregnancy couples. Therefore, a Taegyo program can be recommended as a nursing intervention program for first pregnancy couples.