RESUMO
Purpose: Bone repair aims to restore the anatomical, biomechanical, and functional integrity of the affected structure. Here we study the effects of ascorbic acid (AA) and epidermal growth factor (EGF) applied in a single dose and in combination on the repair of a noncritical bone defect model. Methods: Twenty-four rats were divided into four groups: an intact G-1 control group, and three groups that underwent a noncritical bone defect in the right tibia: G-2 treated with AA, G-3 treated with EGF, and G-4 treated with AA in combination with EGF. After 21 days of treatment, rats were sacrificed, the tibias were dissected and a destructive biomechanical analysis of three-point flexion test was performed in a universal testing machine; the values of stiffness, resistance, maximum energy, and energy at maximum load were statistically compared. Results: G-3 and G-4 recovered the biomechanical properties of strength and stiffness of an intact tibia 3 weeks after their application. Not so the energy and energy at maximum load. For G-2, only the stiffness of an intact tibia was recovered. Conclusion: EGF and AA-EGF applied to a noncritical bone defect in the rat tibia favors the recovery of bone resistance and stiffness.
Assuntos
Animais , Ratos , Ácido Ascórbico/análise , Tíbia/cirurgia , Materiais Biocompatíveis/análise , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fenômenos Biomecânicos , Procedimentos Ortopédicos/métodosRESUMO
To investigate the influence of Kuntai capsules on the expression level of leukemia inhibitory factor (LIF), insulin-like growth factor-I (IGF-1)and epidermal growth factor (EGF) during the mouse's implantation window of superovulation period and controlled ovarian hyperstimulation period. 90 female mice were randomly divided into six groups in control, superovulation and controlled ovarian hyperstimulation (COH) conditions. The RNA expression of EGF, LIF and IGF-1 in the endometrium on the 4th day of pregnancy was detected, and the relative expression was compared. mRNA expression of these three factors in endometrium was significantly lower in superovulation and COH groups than control group (p<0.001). mRNA expression of these three factors in endometrium remained obviously lower in superovulation plus kuntai capsule group and COH plus kuntai capsule group than control group (p<0.01). mRNA expression of these three factors in endometrium was lower in control group than in the NS plus kuntai capsule group (p<0.05). Kuntai capsule cannot completely reverse the endometrial damages caused by superovulation and COH. Thus Kuntai capsule could partially improve a mouse's endometrial receptivity during the implantation window.
Para investigar la influencia de las cápsulas de Kuntai en el nivel de expresión del factor inhibidor de la leucemia (LIF), el factor de crecimiento similar a la insulina I (IGF-1) y el factor de crecimiento epidérmico (EGF) durante la ventana de implantación del ratón del período de superovulación y la hiperestimulación ovárica controlada período, se dividieron aleatoriamente 90 ratones hembra en seis grupos en condiciones de control, superovulación e hiperestimulación ovárica controlada (COH). Se detectó la expresión de ARN de EGF, LIF e IGF-1en el endometrio al cuarto día de embarazo, y se comparó la expresión relativa. La expresión de ARNm de estos tres factores en el endometrio fue significativamente menor en los grupos de superovulación y COH que en el grupo control (p<0,001). La expresión de ARNm de estos tres factores en el endometrio permaneció más baja en el grupo de cápsulas de superovulación más Kuntai y en el grupo de cápsulas de COH más Kuntai respecto del grupo control (p<0,01). La expresión de ARNm de estos tres factores en el endometrio fue menor en el grupo control que en el grupo de cápsula NS más Kuntai (p<0,05). La cápsula de Kuntai no pudo revertir completamente los daños endometriales causados por la superovulación y la COH. Por lo tanto, se sugiere que la cápsula de Kuntai podría mejorar parcialmente la receptividad endometrial de un ratón durante la ventana de implantación.
Assuntos
Animais , Feminino , Camundongos , Indução da Ovulação/métodos , Somatomedinas/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator Inibidor de Leucemia/efeitos dos fármacos , Implantação do Embrião , Superovulação , Somatomedinas/genética , Somatomedinas/metabolismo , Cápsulas , Reação em Cadeia da Polimerase/métodos , Eletroforese , Fator de Crescimento Epidérmico/genética , Fator de Crescimento Epidérmico/metabolismo , Fator Inibidor de Leucemia/genética , Fator Inibidor de Leucemia/metabolismoRESUMO
Erlotinib is a widely used, reversible tyrosine kinase inhibitor (TKI), targeting pro-proliferative signaling of epidermal growth factor receptor (EGFR). The drug is approved for the first-line treatment of patients with metastatic non-small cell lung cancer with EGFR mutations. Extracellular glycans can affect EGFR expression, dimerization, phosphorylation, and EGF binding. In this study we investigated the effects of EGF and erlotinib on the cell cycle of naive and sialidase (alpha-neuraminidase)-pretreated human A549 alveolar epithelial cells. A549 cells were labeled with propidium iodide, and fractions of cells in different phases of cycle were quantified by flow cytometry. We found that neither did desialilation nor EGF, as well as erlotinib treatment, increase the number of damaged cells (subG0/G1 cell fraction), while erlotinib did significantly increase the number of G0/G1 cells and decrease S + G2/M cell fractions. In naive cells, EGF increased proliferating cell numbers by more than 40%, and this effect was blocked by erlotinib. In desialylated cells, however, proliferation was significantly decreased by about 29%, and EGF and erlotinib did not exert significant effects. We conclude that changes in alveolar epithelial cell membrane glycosylation may affect function of growth-promoting receptors and erlotinib effectiveness.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Fator de Crescimento Epidérmico , Cloridrato de Erlotinib , Neoplasias Pulmonares , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Fator de Crescimento Epidérmico/efeitos dos fármacos , Células Epiteliais , Cloridrato de Erlotinib/farmacologia , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Neuraminidase , Inibidores de Proteínas Quinases , QuinazolinasRESUMO
PURPOSE:: To investigate the effect of nicotinamide on the secretion of pro-an giogenic and pro-inflammatory cytokines in uveal melanoma cell lines. METHODS:: Two human uveal melanoma cell lines (92.1 and OCM-1) were treated with nicotinamide (10 mmol/L) or control media for 48 hours in culture. The su perna tant from each culture was used in sandwich enzyme-linked immuno sorbent assay-based angiogenesis and inflammation arrays to evaluate the effects of exogenously administered nicotinamide on the secretion of a total of 20 pro-an gio genic and pro-inflammatory proteins. RESULTS:: Seven pro-angiogenic cytokines were detected under control conditions for both uveal melanoma cell lines. Treatment with nicotinamide resulted in a significant decrease in secretion of the following pro-angiogenic cytokines: angiogenin, angiopoietin-2, epidermal growth factor, and vascular epithelial growth factor-A in the 92.1 cells; basic fibroblast growth factor in the OCM-1 cells; and placenta growth factor in both cell lines. Among the pro-inflammatory proteins, monocyte chemotactic protein-1 and interleukin-8 were expressed in both untreated cell lines and both were significantly reduced when treated with nicotinamide. CONCLUSIONS:: Results from this in vitro model suggest that nicotinamide may have anti-inflammatory and anti-angiogenic properties, which may open the possibility of using it as a chemopreventive agent for uveal melanoma; however, further studies including animal models are warranted.
Assuntos
Inibidores da Angiogênese/farmacologia , Anti-Inflamatórios/farmacologia , Citocinas/efeitos dos fármacos , Melanoma/metabolismo , Niacinamida/farmacologia , Neoplasias Uveais/metabolismo , Angiopoietina-2/metabolismo , Linhagem Celular Tumoral , Quimiocina CCL2/efeitos dos fármacos , Citocinas/metabolismo , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/efeitos dos fármacos , Humanos , Interleucina-8/efeitos dos fármacos , Melanoma/irrigação sanguínea , Fator de Crescimento Placentário/efeitos dos fármacos , Ribonuclease Pancreático/efeitos dos fármacos , Neoplasias Uveais/irrigação sanguíneaRESUMO
ABSTRACT Purpose: To investigate the effect of nicotinamide on the secretion of pro-an giogenic and pro-inflammatory cytokines in uveal melanoma cell lines. Methods: Two human uveal melanoma cell lines (92.1 and OCM-1) were treated with nicotinamide (10 mmol/L) or control media for 48 hours in culture. The su perna tant from each culture was used in sandwich enzyme-linked immuno sorbent assay-based angiogenesis and inflammation arrays to evaluate the effects of exogenously administered nicotinamide on the secretion of a total of 20 pro-an gio genic and pro-inflammatory proteins. Results: Seven pro-angiogenic cytokines were detected under control conditions for both uveal melanoma cell lines. Treatment with nicotinamide resulted in a significant decrease in secretion of the following pro-angiogenic cytokines: angiogenin, angiopoietin-2, epidermal growth factor, and vascular epithelial growth factor-A in the 92.1 cells; basic fibroblast growth factor in the OCM-1 cells; and placenta growth factor in both cell lines. Among the pro-inflammatory proteins, monocyte chemotactic protein-1 and interleukin-8 were expressed in both untreated cell lines and both were significantly reduced when treated with nicotinamide. Conclusions: Results from this in vitro model suggest that nicotinamide may have anti-inflammatory and anti-angiogenic properties, which may open the possibility of using it as a chemopreventive agent for uveal melanoma; however, further studies including animal models are warranted.
RESUMO Objetivo: Acredita-se que a nicotinamida (NIC) seja capaz de diminuir a angiogênese induzida pelo fator de crescimento endotelial vascular (VEGF). Investigar os efeitos da nicotinamida sobre a secreção de citocinas pró-angiogênicas e pró-inflamatórias em linhagens de células de melanoma uveal humano (UM). Métodos: Duas linhagens de células humanas de UM (92,1 e OCM-1) foram tratadas com NIC (10 mmol/L) ou apenas com meio de cultura por 48 horas. O sobrenadante das culturas obtido após a administração de nicotinamida foi comparado com o sobrenadante das culturas controle quanto à expressão de 20 fatores pró-angiogênicos e pró-inflamatórios, pela técnica de enzyme-linked immunosorbent assay (ELISA). Resultados: Sete citocinas pró-angiogênicas foram detectadas nas condições de controle em ambas as linhagens de células de UM. O tratamento com nicotinamida promoveu uma redução significativa da secreção das seguintes citocinas angiogênicas: Angiogenina, ANG2, EGF e VEGF-A em células 92.1; bFGF em células OCM-1; PIGF em ambas as linhagens celulares. Quanto às proteínas pró-inflamatórias, a expressão de MCP-1 e IL-8 foi significativamente reduzida com a administração de nicotinamida em relação às culturas de células que não receberam o tratamento. Conclusões: Nicotinamida apresenta propriedades anti-inflamatórias e anti-angiogênicas em modelo experimental in vitro. Tais efeitos sugerem a possibilidade de utilizar esta substância na quimioprevenção do UM. Entretanto, estudos com modelos experimentais in vivo são necessários para melhor avaliar o benefício do tratamento do UM com nicotinamida.
Assuntos
Humanos , Neoplasias Uveais/metabolismo , Citocinas/efeitos dos fármacos , Niacinamida/farmacologia , Inibidores da Angiogênese/farmacologia , Melanoma/metabolismo , Anti-Inflamatórios/farmacologia , Ribonuclease Pancreático/efeitos dos fármacos , Neoplasias Uveais/irrigação sanguínea , Citocinas/metabolismo , Fator 2 de Crescimento de Fibroblastos/efeitos dos fármacos , Interleucina-8/efeitos dos fármacos , Quimiocina CCL2/efeitos dos fármacos , Linhagem Celular Tumoral , Angiopoietina-2/metabolismo , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator de Crescimento Placentário/efeitos dos fármacos , Melanoma/irrigação sanguíneaRESUMO
Betel quid (BQ) chewing is an etiologic factor of oral submucous fibrosis (OSF) and oral cancer. There are 600 million BQ chewers worldwide. The mechanisms for the toxic and inflammatory responses of BQ are unclear. In this study, both areca nut (AN) extract (ANE) and arecoline stimulated epidermal growth factor (EGF) and interleukin-1α (IL-1α) production of gingival keratinocytes (GKs), whereas only ANE can stimulate a disintegrin and metalloproteinase 17 (ADAM17), prostaglandin E2 (PGE2) and 8-isoprostane production. ANE-induced EGF production was inhibited by catalase. Addition of anti-EGF neutralizing antibody attenuated ANE-induced cyclooxygenase-2 (COX-2), mature ADAM9 expression and PGE2 and 8-isoprostane production. ANE-induced IL-1α production was inhibited by catalase, anti-EGF antibody, PD153035 (EGF receptor antagonist) and U0126 (MEK inhibitor) but not by α-naphthoflavone (cytochrome p450-1A1 inhibitor). ANE-induced ADAM17 production was inhibited by pp2 (Src inhibitor), U0126, α-naphthoflavone and aspirin. AG490 (JAK inhibitor) prevented ANE-stimulated ADAM17, IL-1α, PGE2 production, COX-2 expression, ADAM9 maturation, and the ANE-induced decline in keratin 5 and 14, but showed little effect on cdc2 expression and EGF production. Moreover, ANE-induced 8-isoprostane production by GKs was inhibited by catalase, anti-EGF antibody, AG490, pp2, U0126, α-naphthoflavone, Zinc protoporphyrin (ZnPP) and aspirin. These results indicate that AN components may involve in BQ-induced oral cancer by induction of reactive oxygen species, EGF/EGFR, IL-1α, ADAMs, JAK, Src, MEK/ERK, CYP1A1, and COX signaling pathways, and the aberration of cell cycle and differentiation. Various blockers against ROS, EGF, IL-1α, ADAM, JAK, Src, MEK, CYP1A1, and COX can be used for prevention or treatment of BQ chewing-related diseases.
Assuntos
Areca/toxicidade , Gengiva/efeitos dos fármacos , Queratinócitos/efeitos dos fármacos , Extratos Vegetais/toxicidade , Transdução de Sinais/efeitos dos fármacos , Proteína ADAM17/efeitos dos fármacos , Proteína ADAM17/metabolismo , Linhagem Celular , Dinoprosta/análogos & derivados , Dinoprosta/metabolismo , Dinoprostona/metabolismo , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator de Crescimento Epidérmico/metabolismo , Humanos , Interleucina-1alfa/metabolismo , Janus Quinases/efeitos dos fármacos , Janus Quinases/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Objective To observe the effects of Hedyotis diffusa extract (HDE) on the prolifera- tion, apoptosis, and inflammatory factors of HaCaT cells, and to explore its possible molecular mecha- nisms. Methods HaCaT cells were divided into 3 groups, the vehicle group, the control group, and 3 dose HDE groups. No epidermal growth factor (EGF) or HDE was added in the vehicle group. EGF was added with no HDE treatment in the control group. HDE (25, 50, 100 mg/mL) and EGF were added in the 3 HDE groups to co-culture HaCaT cells. The effects of HDE on EGF-induced proliferation of HaCaT cells were de- tected using CCK-8 assay. The effects of HDE on the growth cycle and apoptosis rate of HaCaT cells were measured using flow cytometry. Moreover, protein expression levels of Ki67, Bcl-xL, clAP1 , procaspase- 3, and cleaved caspase-3 were determined using Western blot. In addition, levels of IL-6, IL-8, and IL-10 in the supernatant were detected using ELISA. The level of phosphoration of NF-κB p65 (p-p65) was meas- ured using Western blot. Results Compared with the vehicle group, the absorbance of HaCaT cells and the expression level of Ki67 increased (P <0. 05, P <0. 01) ; levels of p-p65, IL-6, and IL-8 were elevated (P <0. 05, P <0. 01); IL-10 level was lowered (P <0.01) in the control group. Compared with the control group, the absorbance of HaCaT cells and the expression level of Ki67 decreased (P <0.05, P <0.01) ; levels of p-p65, IL-6, and IL-8 were reduced (P <0. 05, P <0. 01); IL-10 level was elevated (P <0. 05, P < 0.01) in the 3 dose HDE groups. Meanwhile, the apoptosis rate of HaCaT cells increased more in the 3 dose HDE groups than in the control group (P <0. 05, P <0. 01). The percentage of HaCaT cells at G1 phase was 58. 51 %, 73.12%, and 79. 95% in 25, 50, and 100 mg/mL HDE groups, respectively, showing statisti- cal difference when compared with that in the control group (52. 85%; P <0. 05, P <0. 01). The percentage and apoptosis rate of HaCaT cells at G1 phase were elevated more in 50 and 100 mg/mL HDE groups than in 25 mg/mL HDE group (P <0. 01). Besides, the percentage and apoptosis rate of HaCaT cells at G1 phase were elevated more in 100 mg/mL HDE group than in 50 mg/mL HDE group (P <0. 05). Compared with the control group, protein expression levels of Bcl-xL and cIAP1 were reduced in 100 mg/mL HDE group (P < 0. 01). There was no statistical difference in caspase-3 total amount (P >0. 05), but cleaved caspase-3 ex- pression increased with statistical difference (P <0. 01). Conclusion HDE inhibited the proliferation of HaCaT cells possibly by arresting HaCaT cell growth at G1 phase, promoted the apoptosis of HaCaT cells by stressing protein expressions of Bcl-xL and cIAPI , and elevating protein expressions of cleaved caspase-3, and suppressed inflammatory response of HaCaT cells via regulating NF-κB expression.
Assuntos
Fator de Crescimento Epidérmico , Hedyotis , Extratos Vegetais , Fator de Necrose Tumoral alfa , Apoptose/efeitos dos fármacos , Proliferação de Células , Células Cultivadas , Fator de Crescimento Epidérmico/efeitos dos fármacos , Hedyotis/química , Inflamação , Extratos Vegetais/farmacologia , Fator de Necrose Tumoral alfa/efeitos dos fármacosRESUMO
We previously demonstrated that non-toxic doses of Celecoxib induced the immediate phosphorylation of Erk1-2 in colon tumor associated fibroblasts (TAFs), increasing their responsiveness to epidermal growth factor (EGF). We have now identified two concomitant mechanisms explaining the EGF-Celecoxib cooperation. We found that a 24-48h Celecoxib priming increased EGF receptor (EGFR) mRNA and protein levels in colon TAFs, promoting EGF binding and internalization. Celecoxib-primed TAFs showed a reduced EGFR degradation after EGF challenge. This delay corresponded to a deferred dissociation of EEA1 from EGFR positive endosomes and the accumulation of Rab7, pro Cathepsin-D and SQSTM1/p62, suggesting a shared bottleneck in the pathways of late-endosomes/autophagosomes maturation. Celecoxib modulated the levels of target proteins similarly to the inhibitors of endosome/lysosome acidification Bafilomycin-A1 and NH(4)Cl. Cytoplasmic vesicles fractionation showed a reduced maturation of Cathepsin-D in late endosomes and an increased content of EGFR and Rab7 in lysosomes of Celecoxib-treated TAFs.Our data indicate a double mechanism mediating the increased response to EGF of colon TAFs treated with Celecoxib. While EGFR overexpression could be targeted using anti EGFR drugs, the effects on endosome trafficking and protein turnover represents a more elusive target and should be taken into account for any long-term therapy with Celecoxib.
Assuntos
Celecoxib/farmacologia , Neoplasias do Colo/metabolismo , Inibidores de Ciclo-Oxigenase 2/farmacologia , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB/metabolismo , Fibroblastos/efeitos dos fármacos , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/patologia , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fibroblastos/metabolismo , Citometria de Fluxo , Imunofluorescência , Humanos , Transporte Proteico/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacosRESUMO
Berberine is a wellknown component of the Chinese herbal medicine Huanglian (Coptis chinensis), and is capable of inhibiting the proliferation of multiple cancer cell lines. However, information available regarding the effect of berberine on prostate cancer cell growth is limited. In the present study, LnCaP and PC3 human prostate cancer cell lines were selected as in vitro models in order to assess the efficacy of berberine as an anticancer agent. A cell proliferation assay demonstrated that berberine inhibited cell growth in a doseand timedependent manner. Further investigation revealed berberine significantly accumulated inside cells that were in the G1 phase of the cell cycle and enhanced apoptosis. Western blot analysis demonstrated that berberine inhibited the expression of prostatespecific antigen and the activation of epidermal growth factor receptor (EGFR), and it attenuated EGFR activation following EGF treatment in vitro. In conclusion, the results indicate that berberine inhibits the proliferation of prostate cancer cells through apoptosis and/or cell cycle arrest by inactivation of the EGFR signaling pathway.
Assuntos
Berberina/farmacologia , Receptores ErbB/metabolismo , Neoplasias da Próstata/metabolismo , Transdução de Sinais/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator de Crescimento Epidérmico/metabolismo , Expressão Gênica , Humanos , Masculino , Antígeno Prostático Específico/genética , Antígeno Prostático Específico/metabolismo , Neoplasias da Próstata/genéticaRESUMO
The effects of thallium [Tl(I) and Tl(III)] on the PC12 cell cycle were evaluated without (EGF(-)) or with (EGF(+)) media supplementation with epidermal growth factor (EGF). The following markers of cell-cycle phases were analyzed: cyclin D1 (G1 ); E2F-1, cyclin E and cytosolic p21 (G1 âS transition); nuclear PCNA and cyclin A (S); and cyclin B1 (G2). The amount of cells in each phase and the activation of the signaling cascade triggered by EGF were also analyzed. Tl(I) and Tl(III) (5-100 µM) caused dissimilar effects on PC12 cell proliferation. In EGF(-) cells, Tl(I) increased the expression of G1 âS transition markers and nuclear PCNA, without affecting cyclin A or cyclin B1. In addition to those, cyclin B1 was also increased in EGF(+) cells. In EGF(-) cells, Tl(III) increased the expression of cyclin D1, all the G1âS and S phase markers and cyclin B1. In EGF(+) cells, Tl(III) increased cyclin D1 expression and decreased all the markers of G1 âS transition and the S phase. Even when these cations did not induce the activation of EGF receptor (EGFR) in EGF(-) cells, they promoted the phosphorylation of ERK1/2 and Akt. In the presence of EGF, the cations anticipated EGFR phosphorylation without affecting the kinetics of EGF-dependent ERK1/2 and Akt phosphorylation. Altogether, results indicate that Tl(I) promoted cell proliferation in both EGF(-) and EGF(+) cells. In contrast, Tl(III) promoted the proliferation of EGF(-) cells but delayed it in EGF(+) cells, which may be related to the toxic effects of this cation in PC12 cells.
Assuntos
Ciclo Celular/efeitos dos fármacos , Ciclinas/efeitos dos fármacos , Fator de Crescimento Epidérmico/efeitos dos fármacos , Titânio/toxicidade , Animais , Cátions , Proteínas de Ciclo Celular/efeitos dos fármacos , Proteínas de Ciclo Celular/genética , Proliferação de Células/efeitos dos fármacos , Receptores ErbB/efeitos dos fármacos , Receptores ErbB/genética , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteína Oncogênica v-akt/biossíntese , Proteína Oncogênica v-akt/genética , Oxirredução , Células PC12 , Fosforilação , Ratos , Transdução de Sinais/efeitos dos fármacosRESUMO
Acinar cell regeneration from tubular structures has been reported to occur in duct-deligated salivary glands. However, the detailed process of acinar cell regeneration has not been clarified. We have developed a mouse duct ligation model to clarify the mechanisms underlying acinar cell regeneration, and we analyzed the epidermal growth factor receptor (EGFR) and epidermal growth factor (EGF) ligands using the model. We studied these ligands expressions in the course of acinar cell regeneration using immunohistochemistry and RT-PCR methods. In the duct-ligated portion of the submandibular gland (SMG) that underwent atrophy, newly formed acinar cells were observed arising from the tubular structures after the release of the duct obstruction. The constitutive expression of EGFR was observed by immunohistochemistry in both the duct-ligated and duct-deligated animals as well as in normal controls. The EGFR phosphorylation detected on the tubular structures after duct ligation paralleled the acinar cell regeneration. RT-PCR showed an increase in the epiregulin and heparin-binding EGF levels from day 0 to day 3 after the release of the duct obstruction. The EGF level was increased only after day 7. In vitro, cultured cells isolated from ligated SMGs proliferated and produced EGF ligands following the addition of epiregulin to the culture medium. These findings suggest that the tubular structures localized in an atrophic gland are the source of acinar cell regeneration of the salivary gland. The induction of EGF ligands, in particular epiregulin, may play an important role in acinar cell regeneration in this model.
Assuntos
Células Acinares/fisiologia , Epirregulina/análise , Regeneração/fisiologia , Ductos Salivares/metabolismo , Doenças da Glândula Submandibular/metabolismo , Glândula Submandibular/metabolismo , Anfirregulina , Animais , Atrofia , Betacelulina/análise , Técnicas de Cultura de Células , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Família de Proteínas EGF/análise , Fator de Crescimento Epidérmico/análise , Fator de Crescimento Epidérmico/efeitos dos fármacos , Epigen/análise , Epirregulina/farmacologia , Receptores ErbB/análise , Receptores ErbB/efeitos dos fármacos , Feminino , Fator de Crescimento Semelhante a EGF de Ligação à Heparina/análise , Calicreínas/análise , Calicreínas/efeitos dos fármacos , Ligadura , Camundongos , Camundongos Endogâmicos C57BL , Peptidilprolil Isomerase/análise , Antígeno Nuclear de Célula em Proliferação/análise , Ductos Salivares/efeitos dos fármacos , Ductos Salivares/patologia , Glândula Submandibular/patologia , Doenças da Glândula Submandibular/patologia , Fator de Crescimento Transformador alfa/análise , Fator de Crescimento Transformador alfa/efeitos dos fármacosRESUMO
PURPOSE: The purpose of this study was to evaluate changes in tear volume, epidermal growth factor (EGF), and histology of the rabbit lacrimal gland after intraglandular application of botulinum toxin. METHODS: Ten New Zealand rabbits were separated into 3 groups. Eight rabbits received botulinum toxin injection (2.5 U per 0.1 mL) into the right lacrimal gland and saline injection (0.1 mL) into the left lacrimal gland as a sham control. Two rabbits were untreated to serve as normal controls. Tear volume was measured using cotton thread every 2 weeks, and 4 rabbits were killed after 2 and 4 weeks. The lacrimal glands were surgically excised and sectioned or lysed for gene expression analysis. Epidermal growth factor expression and concentration were evaluated by real-time polymerase chain reaction and enzyme-linked immunosorbent assay; morphometric and histologic analyses were performed. RESULTS: The rabbits that were killed 2 weeks after the injection showed decreased tear volume and increased EGF expression and concentration, but differences were not statistically significant. The rabbits that were killed 4 weeks after the injection exhibited similar results. When all 8 rabbits were considered, we detected a significant decrease in tear volume and increased EGF expression and concentration (P = 0.012, P = 0.011, and P = 0.012, respectively). The EGF level was not significantly correlated with the tear volume. There were no prominent histologic changes between the glands, and the lumen versus fibrosis ratio in the interlobular ducts showed no statistically significant difference. CONCLUSIONS: The use of botulinum toxin in patients with epiphora is effective, safe, and repeatable because it reduces tear volume and increases the EGF level to prevent corneal damage while causing no histologic changes.
Assuntos
Inibidores da Liberação da Acetilcolina/farmacologia , Toxinas Botulínicas/farmacologia , Aparelho Lacrimal/efeitos dos fármacos , Animais , Lesões da Córnea/prevenção & controle , Tecido Elástico/efeitos dos fármacos , Tecido Elástico/patologia , Ensaio de Imunoadsorção Enzimática , Fator de Crescimento Epidérmico/efeitos dos fármacos , Perfilação da Expressão Gênica , Aparelho Lacrimal/metabolismo , Aparelho Lacrimal/patologia , Doenças do Aparelho Lacrimal/tratamento farmacológico , Masculino , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Lágrimas/efeitos dos fármacos , Lágrimas/metabolismo , Fatores de TempoRESUMO
BACKGROUND: Ulcerative colitis is a chronic inflammatory disease and involves multiple etiological factors. Acetic acid (AA)-induced colitis is a reproducible and simple model, sharing many characteristics with human colitis. N-acetylcysteine (NAC) has been widely used as an antioxidant in vivo and in vitro. NAC can affect several signaling pathways involving in apoptosis, angiogenesis, cell growth and arrest, redox-regulated gene expression, and inflammatory response. Therefore, NAC may not only protect against the direct injurious effects of oxidants, but also beneficially alter inflammatory events in colitis. This study was conducted to investigate whether NAC could alleviate the AA-induced colitis in a porcine model. METHODS: Weaned piglets were used to investigate the effects of NAC on AA-induced colitis. Severity of colitis was evaluated by colon histomorphology measurements, histopathology scores, tissue myeloperoxidase activity, as well as concentrations of malondialdehyde and pro-inflammatory mediators in the plasma and colon. The protective role of NAC was assessed by measurements of antioxidant status, growth modulator, cell apoptosis, and tight junction proteins. Abundances of caspase-3 and claudin-1 proteins in colonic mucosae were determined by the Western blot method. Epidermal growth factor receptor, amphiregulin, tumor necrosis factor-alpha (TNF-α), and toll-like receptor 4 (TLR4) mRNA levels in colonic mucosae were quantified using the real-time fluorescent quantitative PCR. RESULTS: Compared with the control group, AA treatment increased (P < 0.05) the histopathology scores, intraepithelial lymphocyte (IEL) numbers and density in the colon, myeloperoxidase activity, the concentrations of malondialdehyde and pro-inflammatory mediators in the plasma and colon, while reducing (P < 0.05) goblet cell numbers and the protein/DNA ratio in the colonic mucosa. These adverse effects of AA were partially ameliorated (P < 0.05) by dietary supplementation with NAC. In addition, NAC prevented the AA-induced increase in caspase-3 protein, while stimulating claudin-1 protein expression in the colonic mucosa. Moreover, NAC enhanced mRNA levels for epidermal growth factor and amphiregulin in the colonic mucosa. CONCLUSION: Dietary supplementation with NAC can alleviate AA-induced colitis in a porcine model through regulating anti-oxidative responses, cell apoptosis, and EGF gene expression.
Assuntos
Ácido Acético , Acetilcisteína/farmacologia , Colite Ulcerativa , Colite/prevenção & controle , Sequestradores de Radicais Livres/farmacologia , Acetilcisteína/uso terapêutico , Anfirregulina , Animais , Apoptose/efeitos dos fármacos , Caspase 3/efeitos dos fármacos , Caspase 3/metabolismo , Claudina-1/efeitos dos fármacos , Claudina-1/metabolismo , Colite/induzido quimicamente , Colite/patologia , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Suplementos Nutricionais , Dinoprostona/metabolismo , Modelos Animais de Doenças , Família de Proteínas EGF , Fator de Crescimento Epidérmico/sangue , Fator de Crescimento Epidérmico/efeitos dos fármacos , Receptores ErbB/efeitos dos fármacos , Receptores ErbB/genética , Sequestradores de Radicais Livres/uso terapêutico , Glicoproteínas/efeitos dos fármacos , Glicoproteínas/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Interleucina-6/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Suínos , Receptor 4 Toll-Like/efeitos dos fármacos , Receptor 4 Toll-Like/genética , Fator de Crescimento Transformador alfa/efeitos dos fármacos , Fator de Crescimento Transformador alfa/metabolismo , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Since centuries, natural compounds from plants, animals and microorganisms were used in medicinal traditions to treat various diseases without a solid scientific basis. Recent studies have shown that plants that were used or are still used in the medieval European medicine are able to provide relieve for many diseases including cancer. Here we summarize impact and effect of selected purified active natural compounds from plants used in European medieval medicinal traditions on cancer hallmarks and enabling characteristics identified by Hanahan and Weinberg. The aim of this commentary is to discuss the pharmacological effect of pure compounds originally discovered in plants with therapeutic medieval use. Whereas many reviews deal with Ayurvedic traditions and traditional Chinese medicine, to our knowledge, the molecular basis of European medieval medicinal approaches are much less documented.
Assuntos
Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Neoplasias/tratamento farmacológico , Plantas Medicinais/química , Animais , Ciclo Celular/efeitos dos fármacos , Fator de Crescimento Epidérmico/efeitos dos fármacos , Europa (Continente) , Humanos , Sistema Imunitário/efeitos dos fármacos , Terapia de Alvo Molecular/métodos , Neoplasias/patologia , Neovascularização Patológica/tratamento farmacológico , FitoterapiaRESUMO
BACKGROUND: Tacrolimus ointment has shown efficacy in treating T-cell-mediated inflammatory oral mucosal diseases, including lichen planus. However, the safety of topical tacrolimus has been questioned, based on its possible association with malignant transformation. AIM: To evaluate the safety aspects of tacrolimus in a three-dimensional in vitro model of oral mucosa containing both multilayered epithelium and connective tissue (raft culture). METHODS: Raft cultures mimicking oral mucosa were topically exposed to tacrolimus, and the effects on cell proliferation and adhesion, epidermal growth factor receptors (EGFR, ERBB2, ERBB3, ERBB4), and apoptosis were evaluated with immunohistochemistry and terminal dUTP nick-end labelling, respectively. Results. The epithelium of the cultures was found to be slightly thinner, but no changes in cell proliferation or adhesion, apoptosis, or expression of epidermal growth factor receptors were detected. CONCLUSIONS: Our results suggest that short-term topical tacrolimus exposure of in vitro constructed oral mucosa does not induce changes in a number of factors known to be involved in malignant transformation.
Assuntos
Imunossupressores/farmacologia , Mucosa Bucal/efeitos dos fármacos , Tacrolimo/farmacologia , Apoptose/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Técnicas de Cultura de Células/métodos , Proliferação de Células/efeitos dos fármacos , Fator de Crescimento Epidérmico/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Imunossupressores/efeitos adversos , Marcação In Situ das Extremidades Cortadas/métodos , Tacrolimo/efeitos adversos , Células Tumorais CultivadasRESUMO
Piceatannol (trans-3,4,3',5'-tetrahydroxystilbene) is a polyphenol detected in grapes, red wine and Rheum undulatum; it has also been demonstrated to exert anticarcinogenic effects. In this study, in order to determine whether piceatannol inhibits the lung metastasis of prostate cancer cells, MAT-Ly-Lu (MLL) rat prostate cancer cells expressing luciferase were injected into the tail veins of male nude mice. The oral administration of piceatannol (20 mg/kg) significantly inhibited the accumulation of MLL cells in the lungs of these mice. In the cell culture studies, piceatannol was demonstrated to inhibit the basal and epidermal growth factor (EGF)-induced migration and invasion of DU145 cells, in addition to the migration of MLL, PC3 and TRAMP-C2 prostate cancer cells. In DU145 cells, piceatannol attenuated the secretion and messenger RNA levels of matrix metalloproteinase-9, urokinase-type plasminogen activator (uPA) and vascular endothelial growth factor (VEGF). Piceatannol increased the protein levels of tissue inhibitor of metalloproteinase-2 in a concentration-dependent fashion. Additionally, piceatannol inhibited the phosphorylation of signal transducer and activator of transcription (STAT) 3. Furthermore, piceatannol effected reductions in both basal and EGF-induced interleukin (IL)-6 secretion. An IL-6 neutralizing antibody inhibited EGF-induced STAT3 phosphorylation and EGF-stimulated migration of DU145 cells. Interleukin-6 treatment was also shown to enhance the secretion of uPA and VEGF, STAT3 phosphorylation and the migration of DU145 cells; these increases were suppressed by piceatannol. These results demonstrate that the inhibition of IL-6/STAT3 signaling may constitute a mechanism by which piceatannol regulates the expression of proteins involved in regulating the migration and invasion of DU145 cells.
Assuntos
Interleucina-6/antagonistas & inibidores , Interleucina-6/metabolismo , Transdução de Sinais/efeitos dos fármacos , Estilbenos/farmacologia , Animais , Linhagem Celular Tumoral , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator de Crescimento Epidérmico/metabolismo , Regulação da Expressão Gênica , Humanos , Masculino , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Metástase Neoplásica/tratamento farmacológico , Metástase Neoplásica/patologia , Fosforilação , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/genética , Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
We report the case of a 63-year-old obese man with a rapid-onset of widespread acanthosis nigricans (AN) in the setting of having recently initiated treatment with niacin for dyslipidemia. Although obesity and insulin-resistance are risk factors for AN, AN associated with endocrine dysfunction tends to have a more gradual onset and limited involvement. Owing to our patient's age, the rapid onset, and extensive distribution of his eruption, we initially were concerned about paraneoplastic AN. However, an evaluation for a malignant condition was negative. The timing of the onset of our patient's eruption within several months of starting niacin therapy is consistent with niacin-induced AN. Niacin is known to cause rapidly progressive, widespread AN that is reversible upon discontinuation of the medication. We discuss the pathogenesis of AN, which is thought to be the final common manifestation of stimulation of different subtypes of tyrosine kinase receptors by various epidermal growth factors.
Assuntos
Acantose Nigricans/induzido quimicamente , Hipolipemiantes/efeitos adversos , Niacina/efeitos adversos , Acantose Nigricans/etiologia , Doenças Cardiovasculares/complicações , Diagnóstico Diferencial , Dislipidemias/complicações , Dislipidemias/tratamento farmacológico , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator de Crescimento Epidérmico/fisiologia , Humanos , Hiperinsulinismo/complicações , Hipolipemiantes/uso terapêutico , Resistência à Insulina , Masculino , Pessoa de Meia-Idade , Niacina/uso terapêutico , Obesidade/complicações , Síndromes Paraneoplásicas/diagnóstico , Receptores Proteína Tirosina Quinases/fisiologiaRESUMO
Hypertension is characterized by a sustained increase in vasoconstriction and attenuated vasodilation in the face of elevated mechanical stress in the blood vessel wall. To adapt to the increased stress, the vascular smooth muscle cell and its surrounding environment undergo structural and functional changes known as vascular remodeling. Multiple mechanisms underlie the remodeling process, including increased expression of humoral factors and their receptors as well as adhesion molecules and their receptors, all of which appear to collaborate and interact in the response to pressure elevation. In this review, we focus on the interactions between integrin signaling pathways and the activation of growth factor receptors in the response to the increased mechanical stress experienced by blood vessels in hypertension.
Assuntos
Endotélio Vascular/efeitos dos fármacos , Fator de Crescimento Epidérmico/efeitos dos fármacos , Receptores ErbB/efeitos dos fármacos , Hipertensão/tratamento farmacológico , Integrinas/fisiologia , Fenômenos Biomecânicos , Canais de Cálcio/efeitos dos fármacos , Matriz Extracelular/efeitos dos fármacos , Humanos , Fator de Crescimento Insulin-Like I/efeitos dos fármacos , Fenômenos Mecânicos , Músculo Liso Vascular/efeitos dos fármacos , Receptor Tipo 1 de Angiotensina/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
AIMS: In surgical pathology, formalin-fixed paraffin-embedded tissues are increasingly being used as a source of DNA and RNA for molecular assays in addition to histopathological evaluation. However, the commonly used formalin fixative is carcinogenic, and its crosslinking impairs DNA and RNA quality. METHODS: The suitability of three new presumably less toxic, crosslinking (F-Solv) and non-crosslinking (FineFIX, RCL2) alcohol-based fixatives was tested for routine molecular pathology in comparison with neutral buffered formalin (NBF) as gold standard. Size ladder PCR, epidermal growth factor receptor sequence analysis, microsatellite instability (MSI), chromogenic (CISH), fluorescence in situ hybridisation (FISH) and qPCR were performed. RESULTS: The alcohol-based non-crosslinking fixatives (FineFIX and RCL2) resulted in a higher DNA yield and quality compared with crosslinking fixatives (NBF and F-Solv). Size ladder PCR resulted in a shorter amplicon size (300 bp) for both crosslinking fixatives compared with the non-crosslinking fixatives (400 bp). All four fixatives were directly applicable for MSI and epidermal growth factor receptor sequence analysis. All fixatives except F-Solv showed clear signals in CISH and FISH. RNA yield and quality were superior after non-crosslinking fixation. qPCR resulted in lower Ct values for RCL2 and FineFIX. CONCLUSION: The alcohol-based non-crosslinking fixatives performed better than crosslinking fixatives with regard to DNA and RNA yield, quality and applicability in molecular diagnostics. Given the higher yield, less starting material may be necessary, thereby increasing the applicability of biopsies for molecular studies.
Assuntos
DNA/efeitos dos fármacos , Fixadores/farmacologia , Formaldeído/farmacologia , RNA/efeitos dos fármacos , Reagentes de Ligações Cruzadas/farmacologia , DNA/metabolismo , Fator de Crescimento Epidérmico/efeitos dos fármacos , Fator de Crescimento Epidérmico/genética , Humanos , Instabilidade de Microssatélites/efeitos dos fármacos , RNA/metabolismo , Fixação de Tecidos/métodosRESUMO
EGFR, as a critical signaling pathway in many human tumors, has become an important target of cancer drug design. Taspine has shown meaningful angiogenesis activity in previous studies. This paper is to investigate the antitumor action of taspine by modulating the EGFR signaling pathway. The study determined the expression of key signaling molecules of EGFR (EGFR, Akt, p-Akt, Erk, and p-Erk) by Western blot and real-time PCR and analyzed their correlations with subsequent reactions. In addition, the cell proliferation, migration, and EGF production were examined by MTT, transwell system, and ELISA. The antitumor activity in vivo was carried out by xenograft in athymic mice. The results showed that taspine could inhibit A431 and Hek293/EGFR cell proliferation and A431 cell migration as well as EGF production. Compared to the negative control, EGFR, Akt, and phosphorylation of Akt were significantly inhibited by taspine treatment in A431 and HEK293/EGFR cells. Consistent with the inhibition of Akt activity, Erk1/2 and its phosphorylation were reduced. Moreover, taspine inhibited A431 xenograft tumor growth. These results suggest that EGFR activated by EGF and its downstream signaling pathways proteins could be downregulated by taspine in a dose-dependent manner. The antitumor mechanism of taspine through the EGFR pathway lies in the ability to inhibit A431 cell proliferation and migration by reducing EGF secretion. This occurs through the repression of EGFR which mediates not only MAPK (Erk1/2) but also Akt signals.