RESUMO
BACKGROUND: Canine mammary tumors (CMTs) in intact female dogs provide a natural model for investigating metastatic human cancers. Our prior research identified elevated expression of Anterior Gradient 2 (AGR2), a protein disulfide isomerase (PDI) primarily found in the endoplasmic reticulum (ER), in CMT tissues, highly associated with CMT progression. We further demonstrated that increased AGR2 expression actively influences the extracellular microenvironment, promoting chemotaxis in CMT cells. Unraveling the underlying mechanisms is crucial for assessing the potential of therapeutically targeting AGR2 as a strategy to inhibit a pro-metastatic microenvironment and impede tumor metastasis. METHODS: To identify the AGR2-modulated secretome, we employed proteomics analysis of the conditioned media (CM) from two CMT cell lines ectopically expressing AGR2, compared with corresponding vector-expressing controls. AGR2-regulated release of 14-3-3ε (gene: YWHAE) and α-actinin 4 (gene: ACTN4) was validated through ectopic expression, knockdown, and knockout of the AGR2 gene in CMT cells. Extracellular vesicles derived from CMT cells were isolated using either differential ultracentrifugation or size exclusion chromatography. The roles of 14-3-3ε and α-actinin 4 in the chemotaxis driven by the AGR2-modulated CM were investigated through gene knockdown, antibody-mediated interference, and recombinant protein supplement. Furthermore, the clinical relevance of the release of 14-3-3ε and α-actinin 4 was assessed using CMT tissue-immersed saline and sera from CMT-afflicted dogs. RESULTS: Proteomics analysis of the AGR2-modulated secretome revealed increased abundance in 14-3-3ε and α-actinin 4. Ectopic expression of AGR2 significantly increased the release of 14-3-3ε and α-actinin 4 in the CM. Conversely, knockdown or knockout of AGR2 expression remarkably reduced their release. Silencing 14-3-3ε or α-actinin 4 expression diminished the chemotaxis driven by AGR2-modulated CM. Furthermore, AGR2 controls the release of 14-3-3ε and α-actinin 4 primarily via non-vesicular routes, responding to the endoplasmic reticulum (ER) stress and autophagy activation. Knockout of AGR2 resulted in increased α-actinin 4 accumulation and impaired 14-3-3ε translocation in autophagosomes. Depletion of extracellular 14-3-3ε or α-actinin 4 reduced the chemotaxis driven by AGR2-modulated CM, whereas supplement with recombinant 14-3-3ε in the CM enhanced the CM-driven chemotaxis. Notably, elevated levels of 14-3-3ε or α-actinin 4 were observed in CMT tissue-immersed saline compared with paired non-tumor samples and in the sera of CMT dogs compared with healthy dogs. CONCLUSION: This study elucidates AGR2's pivotal role in orchestrating unconventional secretion of 14-3-3ε and α-actinin 4 from CMT cells, thereby contributing to paracrine-mediated chemotaxis. The insight into the intricate interplay between AGR2-involved ER stress, autophagy, and unconventional secretion provides a foundation for refining strategies aimed at impeding metastasis in both canine mammary tumors and potentially human cancers.
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Proteínas 14-3-3 , Actinina , Autofagia , Quimiotaxia , Estresse do Retículo Endoplasmático , Neoplasias Mamárias Animais , Mucoproteínas , Animais , Cães , Proteínas 14-3-3/metabolismo , Proteínas 14-3-3/genética , Feminino , Actinina/metabolismo , Actinina/genética , Neoplasias Mamárias Animais/metabolismo , Neoplasias Mamárias Animais/genética , Neoplasias Mamárias Animais/patologia , Linhagem Celular Tumoral , Quimiotaxia/genética , Autofagia/genética , Estresse do Retículo Endoplasmático/genética , Mucoproteínas/genética , Mucoproteínas/metabolismo , Proteínas Oncogênicas/metabolismo , Proteínas Oncogênicas/genéticaRESUMO
INTRODUCTION: The information on the hard- and soft-tissue factors correlated with tooth display after LeFort I osteotomy, especially in the surgery-first approach (SFA), are limited. This study aimed to correlate different parameters with the maxillary incisor display in patients with skeletal Class III malocclusion and those with cleft lip and palate (CLP) in SFA. METHODS: This study consisted of 35 patients with skeletal Class III malocclusion and 32 with cleft deformities who had undergone orthognathic surgery. Pretreatment and posttreatment lateral cephalometric analysis were obtained. Maxillary incisor display was measured in photographs. The intraclass correlation coefficient was used to assess the intraexaminer repeatability. The Student t test was used to compare the maxillary incisor display between 2 groups. Analysis of covariance was performed with pretreatment measurement as covariates, and the important determinants for maxillary incisor display were identified by adjusting the baseline measurements. RESULTS: The mean increase of maxillary advancement at point A was 5.25 mm and 1.28 mm downward movement for skeletal Class III malocclusion, whereas it was 4.59 mm advancement and 2.16 mm downward movement for patients with CLP. The resulting maxillary incisor display was 2.86 mm for skeletal Class III malocclusion and 2.56 mm for patients with CLP. The covariates for maxillary incisor display before intervention was significantly associated with the maxillary incisor display after intervention (P <0.001). However, the interaction effect of groups was not seen (P = 0.933). The horizontal position of A, vertical position of ANS, and upper lip length were the most predictable parameters (P <0.001, P <0.001, P = 0.048, respectively) for maxillary incisor display in both groups. CONCLUSIONS: Horizontal position of point A, vertical position of ANS, and upper lip length are the most important determinants for maxillary incisor display for patients with skeletal Class III malocclusion and those with CLP.
Assuntos
Fenda Labial , Fissura Palatina , Má Oclusão Classe III de Angle , Humanos , Fenda Labial/complicações , Fenda Labial/cirurgia , Incisivo , Fissura Palatina/complicações , Fissura Palatina/cirurgia , Má Oclusão Classe III de Angle/cirurgia , Má Oclusão Classe III de Angle/complicações , Maxila/cirurgiaRESUMO
Few studies have explored the relationship between long-term exposure to particulate matter with an aerodynamic diameter of ≤2.5 µm (PM2.5) and osteoporotic fracture, particularly in high PM2.5 level areas. The aim of this study was to assess the association between long-term exposure to PM2.5 and osteoporotic fracture. We performed a matched case-control study of 16,175 participants obtained from a hospital registry during 2005-2014 in Taiwan. A major osteoporotic fracture was defined as a fracture of the spine, hip, proximal humerus, and forearm. We applied satellite-based spatiotemporal models with 1-km resolution to individually calculate the 1-year average PM2.5 concentration before the index date which was defined as the first visit date for the osteoporotic fracture. Logistic regression models with and without potential confounding factors were used to estimate odds ratios (OR) and 95% confidence intervals (CI) between PM2.5 and osteoporotic fracture, whereas a restricted cubic spline model was used to estimate the dose-response relationship. The sample's median age was 44.7 years (interquartile range: 30.7, 63.1 years). We observed that long-term PM2.5 exposure was associated with osteoporotic fracture, the OR was 1.12 (95% CI: 1.03, 1.22) per 10-µg/m3 increase in PM2.5 in women. In the dose-response association, the OR of osteoporotic fracture was significantly increased for PM2.5 exposures more than 41 µg/m3. We did not find a significant association between PM2.5 (per 10-µg/m3 increase) and osteoporotic fracture among overall population (adjusted OR, 1.02 [95% CI, 0.97 to 1.08]) and men (adjusted OR, 0.94 [95% CI, 0.86 to 1.02]). The results of the stratified analysis showed that women were more sensitive to the adverse impact of PM2.5 that were men, and evidence was obtained of sex-based effect modification (P for interaction = 0.002). Our findings suggest that long-term exposure to PM2.5 is associated with osteoporotic fracture, particularly among women.
Assuntos
Poluentes Atmosféricos , Poluição do Ar , Fraturas por Osteoporose , Adulto , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/toxicidade , Poluição do Ar/análise , Poluição do Ar/estatística & dados numéricos , Estudos de Casos e Controles , Exposição Ambiental/análise , Exposição Ambiental/estatística & dados numéricos , Feminino , Humanos , Masculino , Fraturas por Osteoporose/induzido quimicamente , Fraturas por Osteoporose/epidemiologia , Material Particulado/análise , Material Particulado/toxicidade , Taiwan/epidemiologiaRESUMO
BACKGROUND: Sphingosine kinase 1 (SPHK1) is an enzyme that converts pro-apoptotic ceramide and sphingosine into anti-apoptotic sphingosine-1-phosphate. There is growing evidence that SPHK1 activation promotes oncogenic transformation, tumor growth, chemotherapy resistance, and metastatic spread. High SPHK1 expression has been associated with a poor prognosis in several human cancers. RESULTS: In the present study, the expression level of SPHK1 was examined in feline mammary tumor (FMT) specimens, and the IHC expression level of SPHK1 was associated with the histological grade of FMTs. IHC analysis of 88 FMT cases revealed that the expression level of SPHK1 was upregulated in 53 tumor tissues (60.2%) compared to adjacent mammary tissues. SPHK1 expression in FMTs was significantly associated with histological grade, presence of lymphovascular invasion, and estrogen receptor negativity. Treatment of primary FMT cells with SPHK1 inhibitors reduced cell viability, indicating that SPHK1 acts to promote FMT cell survival. These results indicate that SPHK1 may play an important role in FMTs and may be a therapeutic target in cats with FMT. CONCLUSIONS: SPHK1 over-expression in breast cancer tissues is associated with a poor prognosis in humans. SPHK1 over-expression in more aggressive FMTs provides support for a potential role of SPHK1 inhibitors for the treatment of FMTs. Targeting SPHK1 has potent cytotoxic effects in primary FMT cells. These findings suggest that further examination of the role SPHK1 plays in FMTs will pave the way for the investigation of SPHK1 inhibitors in future clinical applications.
Assuntos
Doenças do Gato/patologia , Neoplasias Mamárias Animais/enzimologia , Neoplasias Mamárias Animais/patologia , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Animais , Vasos Sanguíneos/patologia , Doenças do Gato/enzimologia , Gatos , Feminino , Regulação Neoplásica da Expressão Gênica , Sistema Linfático/patologia , Glândulas Mamárias Animais/enzimologia , Glândulas Mamárias Animais/metabolismo , Neoplasias Mamárias Animais/genética , Neoplasias Mamárias Animais/metabolismo , Invasividade Neoplásica , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Receptores de Estrogênio/genética , Receptores de Estrogênio/metabolismoRESUMO
BACKGROUND: Suture anchor-based fixations of humeral greater tuberosity (GT) fractures have yielded good outcomes in both clinical and biomechanical studies. Be that as it may, the interface contact properties of these fixations have yet to be elaborated. In response, the contact characteristics of two double-row suture anchor fixations for the management of GT fracture were compared. METHODS: Two suture anchor-based fixation techniques, namely the Double-Row Suture Anchor Fixation (DR) and Suture-Bridge Technique (SB), were used to repair humeral GT fractures in 12 fresh-frozen human cadaveric shoulders. A Tekscan pressure sensor placed between the repaired tuberosity and humerus recorded continuous data points directly after repair and for 60 min at set time intervals. The constructs were then cyclically loaded until 100 N, and the shoulders tested at 0°, 30°, and 60° of abduction. Under an applied force, the contact pressure and contact area of the interface were determined. RESULTS: Although both fixation configurations showed decreased contact pressure and area over time, the SB group had higher contact pressure right after fixation and at all time points thereafter. In contrast, the DR group demonstrated significantly more contact pressure and area at each abduction position with the applied load. Nevertheless, contact pressure and area decreased in response to increasing abduction position for both fixation constructs. CONCLUSION: Findings suggest that despite the SB construct having superior interface contact immediately after fixation, the DR construct offered better contact performance at all abduction angles with applied force. LEVEL OF EVIDENCE: Basic Science, Biomechanics.
Assuntos
Fixação Interna de Fraturas/métodos , Fraturas do Ombro/cirurgia , Âncoras de Sutura , Técnicas de Sutura , Fenômenos Biomecânicos , Cadáver , Fixação Interna de Fraturas/instrumentação , Humanos , Úmero/lesões , Úmero/cirurgia , Pessoa de Meia-Idade , Pressão , Manguito Rotador/cirurgia , Ombro/cirurgiaRESUMO
Different edible oils such as lard and soybean oil have been reported to interact with the gut microbiota, affecting host lipid metabolism. However, whether bacteria derived from the environment influence host lipid metabolism remains unclear. This study aimed to clarify the roles of environmental bacteria in host lipid storage and distribution with various edible oils. Gnotobiotic C57BL/6JNarl mice were inoculated with Lysinibacillus xylanilyticus and Paenibacillus azoreducens and then fed either a normal diet (LabDiet 5010, control group) or a diet containing 60% lard (L-group) or soybean oil (S-group) for 18 months. Interestingly, the S-group accumulated massive amounts of white adipose tissue compared to the L- and control groups, while the L-group displayed more hepatic steatosis and fatty droplets than the other groups. The expression of fatty acid synthase (FAS), hydroxymethylglutaryl-coenzyme A reductase (HMGCR), sterol regulatory element-binding protein 2 (SREBP2), and peroxisome proliferator-activated receptor gamma (PPARγ) in the livers of the L-group were markedly elevated compared to the S-group. FAS and PPARγ protein levels were also markedly elevated. However, there were no differences in the expression of the pro-inflammatory cytokines interleukin-6 and tumor necrosis factor-α between the groups. Our results suggest that environmental bacteria may affect host hepatic inflammation and lipid distribution in the presence of high-fat diets, with different effects depending on the fat type consumed.
Assuntos
Dieta Hiperlipídica/efeitos adversos , Fígado Gorduroso/metabolismo , Fígado Gorduroso/microbiologia , Metabolismo dos Lipídeos , Fígado/metabolismo , Fígado/microbiologia , Animais , Bacillaceae/fisiologia , Gorduras na Dieta/efeitos adversos , Gorduras na Dieta/metabolismo , Fígado Gorduroso/patologia , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Paenibacillus/fisiologia , Óleo de Soja/efeitos adversos , Óleo de Soja/metabolismoRESUMO
Immune interferon (IFN), also known as IFN-γ, promotes not only immunomodulation but also antimicrobial and anticancer activity. After IFN-γ binds to the complex of IFN-γ receptor (IFNGR) 1-IFNGR2 and subsequently activates its downstream signaling pathways, IFN-γ immediately causes transcriptional stimulation of a variety of genes that are principally involved in its biological activities. Regarding IFN-γ-dependent immunosurveillance, IFN-γ can directly suppress tumorigenesis and infection and/or can modulate the immunological status in both cancer cells and infected cells. Regarding the anticancer effects of IFN-γ, cancer cells develop strategies to escape from IFN-γ-dependent cancer immunosurveillance. Immune evasion, including the recruitment of immunosuppressive cells, secretion of immunosuppressive factors, and suppression of cytotoxic T lymphocyte responses, is speculated to be elicited by the oncogenic microenvironment. All of these events effectively downregulate IFN-γ-expressing cells and IFN-γ production. In addition to these extrinsic pathways, cancer cells may develop cellular tolerance that manifests as hyporesponsiveness to IFN-γ stimulation. This review discusses the potential escape mechanisms from IFN-γ-dependent immunosurveillance in tumorigenesis.
Assuntos
Interferon gama/imunologia , Neoplasias/imunologia , Evasão Tumoral/imunologia , Microambiente Tumoral/imunologia , Animais , Humanos , Neoplasias/patologiaRESUMO
Helicobacter pylori infection not only induces gastric inflammation but also increases the risk of gastric tumorigenesis. IFN-γ has antimicrobial effects; however, H. pylori infection elevates IFN-γ-mediated gastric inflammation and may suppress IFN-γ signaling as a strategy to avoid immune destruction through an as-yet-unknown mechanism. This study was aimed at investigating the mechanism of H. pylori-induced IFN-γ resistance. Postinfection of viable H. pylori decreased IFN-γ-activated signal transducers and activators of transcription 1 and IFN regulatory factor 1 not only in human gastric epithelial MKN45 and AZ-521 but also in human monocytic U937 cells. H. pylori caused an increase in the C-terminal tyrosine phosphorylation of Src homology-2 domain-containing phosphatase (SHP) 2. Pharmacologically and genetically inhibiting SHP2 reversed H. pylori-induced IFN-γ resistance. In contrast to a clinically isolated H. pylori strain HP238, the cytotoxin-associated gene A (CagA) isogenic mutant strain HP238(CagAm) failed to induce IFN-γ resistance, indicating that CagA regulates this effect. Notably, HP238 and HP238(CagAm) differently caused SHP2 phosphorylation; however, imaging and biochemical analyses demonstrated CagA-mediated membrane-associated binding with phosphorylated SHP2. CagA-independent generation of reactive oxygen species (ROS) contributed to H. pylori-induced SHP2 phosphorylation; however, ROS/SHP2 mediated IFN-γ resistance in a CagA-regulated manner. This finding not only provides an alternative mechanism for how CagA and ROS coregulate SHP2 activation but may also explain their roles in H. pylori-induced IFN-γ resistance.
Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Infecções por Helicobacter/imunologia , Helicobacter pylori/imunologia , Interferon gama/imunologia , Proteína Tirosina Fosfatase não Receptora Tipo 11/imunologia , Animais , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Carcinogênese , Linhagem Celular Tumoral , Mucosa Gástrica/imunologia , Mucosa Gástrica/microbiologia , Humanos , Inflamação/imunologia , Inflamação/microbiologia , Fator Regulador 1 de Interferon/metabolismo , Interferon gama/antagonistas & inibidores , Camundongos , Camundongos Endogâmicos C57BL , Fosforilação , Proteína Tirosina Fosfatase não Receptora Tipo 11/antagonistas & inibidores , Proteína Tirosina Fosfatase não Receptora Tipo 11/genética , Interferência de RNA , RNA Interferente Pequeno , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição STAT1/metabolismo , Células U937RESUMO
Src homology-2 domain-containing phosphatase (SHP) 2, an oncogenic phosphatase, inhibits type II immune interferon (IFN)-γ signaling by subverting signal transducers and activators of transcription 1 tyrosine phosphorylation and activation. For cancer immunoediting, this study aimed to investigate the decrease of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a tumor suppressor protein, leading to cellular impairment of IFN-γ signaling. In comparison with human lung adenocarcinoma A549 cells, the natural PTEN loss in another human lung adenocarcinoma line, PC14PE6/AS2 cells, presents reduced responsiveness in IFN-γ-induced IFN regulatory factor 1 activation and CD54 expression. Artificially silencing PTEN expression in A549 cells also caused cells to be unresponsive to IFN-γ without affecting IFN-γ receptor expression. IFN-γ-induced inhibition of cell proliferation and cytotoxicity were demonstrated in A549 cells but were defective in PC14PE6/AS2 cells and in PTEN-deficient A549 cells. Aberrant activation of SHP2 by ROS was specifically shown in PC14PE6/AS2 cells and PTEN-deficient A549 cells. Inhibiting ROS and SHP2 rescued cellular responses to IFN-γ-induced cytotoxicity and inhibition of cell proliferation in PC14PE6/AS2 cells. These results demonstrate that a decrease in PTEN facilitates ROS/SHP2 signaling, causing lung cancer cells to become unresponsive to IFN-γ.
Assuntos
Adenocarcinoma/metabolismo , Interferon gama/metabolismo , Neoplasias Pulmonares/metabolismo , PTEN Fosfo-Hidrolase/deficiência , Proteína Tirosina Fosfatase não Receptora Tipo 11/metabolismo , Adenocarcinoma/imunologia , Adenocarcinoma/patologia , Adenocarcinoma de Pulmão , Linhagem Celular Tumoral , Proliferação de Células , Ativação Enzimática , Técnicas de Silenciamento de Genes , Humanos , Interferon gama/imunologia , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologia , PTEN Fosfo-Hidrolase/antagonistas & inibidores , PTEN Fosfo-Hidrolase/genética , Interferência de RNA , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Evasão TumoralRESUMO
BACKGROUND: The widespread use of phthalates as plasticizers has raised public health concerns regarding their adverse effects, including an association with cancer. Although animal investigations have suggested an association between phthalate exposure and hepatocellular carcinoma, the mechanisms are unknown. METHODS: The hepatocellular carcinoma cell line Huh7 was treated with benzyl butyl phthalate (BBP), and then analyzed by total internal reflection fluorescence microscopy, confocal microscopy and double immunogold transmission electron microscopy. Following BBP treatment, mRNA levels were measured by RT-PCR, protein levels were measured using western blot, and vascular endothelial growth factor levels were measured by an enzyme-linked immunosorbent assay. Cell migration and invasion assays were evaluated by transwell, and angiogenesis were performed by a tube formation assay. Nude mice were used to investigate metastasis and angiogenesis in vivo. RESULTS: BBP affected hepatocellular carcinoma progression through the aryl hydrocarbon receptor (AhR) and that benzyl butyl phthalate (BBP) stimulated AhR at the cell surface, which then interacted with G proteins and triggered a downstream signaling cascade. BBP activated AhR through a nongenomic action involving G-protein signaling rather than the classical genomic AhR action. BBP treatment promoted cell migration and invasion in vitro and metastasis in vivo via the AhR/Gß/PI3K/Akt/NF-κB pathway. In addition, BBP induced both in vitro and in vivo angiogenesis through the AhR/ERK/VEGF pathway. CONCLUSIONS: These findings suggest a novel nongenomic AhR mechanism involving G-protein signaling induced by phthalates, which contributes to tumor progression of hepatocellular carcinoma.
Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Ácidos Ftálicos/toxicidade , Receptores de Hidrocarboneto Arílico/metabolismo , Transdução de Sinais , Animais , Carcinoma Hepatocelular/metabolismo , Linhagem Celular Tumoral , Genoma , Células HEK293 , Células Hep G2 , Células Endoteliais da Veia Umbilical Humana , Humanos , Neoplasias Hepáticas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Invasividade Neoplásica/patologia , Transplante de Neoplasias , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
CONTEXT: Liver injury can be induced by various hepatotoxicants, including Pseudomonas aeruginosa exotoxin A (PEA). Our previous study indicated that PEA-induced rat hepatotoxicity was T cells and Kupffer cells dependent. Several reports have demonstrated that non-toxic doses of bacterial lipopolysaccharide (LPS) can protect liver against the chemicals-induced toxicity such as acetaminophen and concanavalin-A. OBJECTIVE: This study aimed to investigate the protecting mechanisms of LPS on PEA-induced hepatotoxicity. RESULTS: Rats pretreated with LPS (40 µg/kg, 12 h before PEA admission) significantly decreased animal mortality, serum enzyme (ALT, AST and T-bil) activities, histopathological changes and hepatocytes apoptosis following challenge with PEA. The concentrations of tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ) and interleukin-2 (IL-2) were reduced, but IL-6 and IL-10 were increased in the serum. In addition, prior treatment of these LPS-pretreated rats with gadolinium chloride (GdCl3), a selective Kupffer cell depletion agent, markedly enhanced liver injury after PEA administration. In contrast, the pretreatment of LPS to T-cell deficient athymic nude rats still display significant attenuation of PEA-induced liver injury. This observation further confirmed our hypothesis that LPS ameliorate PEA-hepatotoxicity was through Kupffer cells but not T cells. Moreover, LPS-induced hepatoprotection ability was neutralized by co-treatment with anti-TNF-α antibodies, but not with anti-IFN-γ antibodies. Finally, replacement of LPS with RS-LPS (Rhodobacter sphaeroides LPS), a Toll like receptor-4 (TLR-4) antagonist, resulted in severe hepatotoxicity. CONCLUSION: These results suggested that Kupffer cells, TNF-α and TLR-4 play central mediator roles during the hepatoprotection against PEA-induced hepatotoxicity conferred by LPS.
Assuntos
ADP Ribose Transferases/antagonistas & inibidores , ADP Ribose Transferases/toxicidade , Toxinas Bacterianas/antagonistas & inibidores , Toxinas Bacterianas/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Exotoxinas/antagonistas & inibidores , Exotoxinas/toxicidade , Lipopolissacarídeos/farmacologia , Fígado/efeitos dos fármacos , Fatores de Virulência/antagonistas & inibidores , Fatores de Virulência/toxicidade , Alanina Transaminase/metabolismo , Animais , Apoptose/efeitos dos fármacos , Aspartato Aminotransferases/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/enzimologia , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Interações Medicamentosas , Gadolínio/farmacologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Interferon gama/metabolismo , Interleucinas/metabolismo , Células de Kupffer/efeitos dos fármacos , Células de Kupffer/metabolismo , Fígado/enzimologia , Masculino , Pseudomonas/metabolismo , Ratos , Ratos Nus , Ratos Wistar , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Exotoxina A de Pseudomonas aeruginosaRESUMO
BACKGROUND: Feline mammary carcinoma (FMC) is one of the most prevalent malignancies of female cats. FMC is highly metastatic and thus leads to poor disease outcomes. Among all metastases, liver metastasis occurs in about 25% of FMC patients. However, the mechanism underlying hepatic metastasis of FMC remains largely uncharacterized. RESULTS: Herein, we demonstrate that FMC-derived extracellular vesicles (FMC-EVs) promotes the liver metastasis of FMC by activating hepatic stellate cells (HSCs) to prime a hepatic premetastatic niche (PMN). Moreover, we provide evidence that sphingosine kinase 1 (SK1) delivered by FMC-EV was pivotal for the activation of HSC and the formation of hepatic PMN. Depletion of SK1 impaired cargo sorting in FMC-EV and the EV-potentiated HSC activation, and abolished hepatic colonization of FMC cells. CONCLUSIONS: Taken together, our findings uncover a previously uncharacterized mechanism underlying liver-metastasis of FMC and provide new insights into prognosis and treatment of this feline malignancy.
RESUMO
The epithelial-to-mesenchymal transition (EMT) is involved in cancer metastasis; nevertheless, interferon (IFN)-γ induces anticancer activities by causing cell growth suppression, cytotoxicity, and migration inhibition. Regarding the poor response to exogenously administered IFN-γ as anticancer therapy, it was hypothesized that malignant cells may acquire a means of escaping from IFN-γ immunosurveillance, likely through an EMT-related process. A genomic analysis of human lung cancers revealed a negative link between the EMT and IFN-γ signaling, while compared to human lung adenocarcinoma A549 cells, IFN-γ-hyporesponsive AS2 cells exhibited mesenchymal characteristics. Chemically, physically, and genetically engineered EMT attenuated IFN-γ-induced IFN regulatory factor 1 transactivation. Poststimulation of transforming growth factor-ß induced the EMT and also selectively retarded IFN-γ-responsive gene expression as well as IFN-γ-induced signal transducer and activator of transcription 1 activation, major histocompatibility complex I, and CD54 expression, cell migration/invasion inhibition, and direct/indirect cytotoxicity. Without changes in IFN-γ receptors, excessive oxidative activation of Src homology-2 containing phosphatase 2 (SHP2) in cells undergoing the EMT primarily caused cellular hyporesponsiveness to IFN-γ signaling and cytotoxicity, while combining an SHP2 inhibitor or antioxidant sensitized EMT-associated AS2 and mesenchymal A549 cells to IFN-γ-induced priming effects on tumor necrosis factor-related apoptosis-inducing ligand cytotoxicity. In cell line-derived xenograft models, combined treatment with IFN-γ and an SHP2 inhibitor induced enhanced anticancer activities. These results imply that EMT-associated SHP2 activation inhibits IFN-γ signaling, facilitating lung cancer cell escape from IFN-γ immunosurveillance.
Assuntos
Interferon gama , Neoplasias Pulmonares , Linhagem Celular Tumoral , Movimento Celular/imunologia , Transição Epitelial-Mesenquimal/imunologia , Humanos , Vigilância Imunológica , Interferon gama/imunologia , Interferon gama/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologiaRESUMO
PURPOSE: This study compares 1) progressive dental and skeletal changes, 2) postsurgical stability, and 3) treatment efficacy of patients with skeletal Class III correction with and without presurgical orthodontic treatment. PATIENTS AND METHODS: The study includes 53 patients who underwent orthognathic surgery (OGS) to correct skeletal Class III malocclusion. The patient grouping is based on presurgical orthodontics: surgical-first (SF) approach (n = 18) and modified-conventional (MC) approach (n = 35). This study divides the MC group into 2 groups based on whether patients underwent tooth extraction in the presurgical phase (Ext group) (n = 10) or not (Nxt group) (n = 25). Serial lateral cephalometric film measurements identify skeletal and dental changes before treatment (T1), before OGS (T2), 1 month after OGS (T3), and at completion of treatment (T4). This investigation reviews the medical charts for treatment progress and duration. RESULTS: In the presurgical phase, the Ext group showed mild retraction of the upper incisors and more proclination of the lower incisors than the Nxt group. The skeletal sagittal parameters were similar from T1 to T4 in both the SF and MC groups. In the SF group, SN/U1 decreased 6.2° after surgery and was mildly proclined at T4; in the MC group, upper incisor inclination (SN/U1) increased 1.8° from T1 to T4, being 9.4° greater than that in the SF group at T4. The lower incisor inclination (MP/L1) was similar at T4 in both groups. In the MC group, the MP/L1 was shown to be proclined 4.5° before surgery, retroclined 1.9° after surgery, and further retroclined 4.5° until T4. The relapse rate of the mandibular setback was 14.3% in the SF group and 15.7% in the MC group without significant differences. The percentage of sagittal relapse less than 2 mm was 50% in the SF group and 54% in the MC group. The ratio was greater in the MC group with a relapse between 2 and 4 mm but lesser with a relapse greater than 4 mm. The Ext group showed a 3-month longer treatment duration than the Nxt group. CONCLUSION: The amount of skeletal correction and postsurgical relapse, as well as treatment duration, were no different in Class III OGS patients with or without presurgical orthodontic treatment. The presurgical work of lower incisor proclination returned to an inclination similar to the initial status after completing treatment. The final outcome of patients evidenced no difference in lower incisor inclination, with or without presurgical orthodontics.
Assuntos
Cefalometria/métodos , Má Oclusão Classe II de Angle/cirurgia , Ortodontia Corretiva , Procedimentos Cirúrgicos Ortognáticos/métodos , Adolescente , Adulto , Feminino , Seguimentos , Humanos , Incisivo/patologia , Masculino , Má Oclusão Classe II de Angle/terapia , Mandíbula/patologia , Mandíbula/cirurgia , Maxila/patologia , Maxila/cirurgia , Osso Nasal/patologia , Osteotomia/métodos , Osteotomia de Le Fort/métodos , Planejamento de Assistência ao Paciente , Cuidados Pré-Operatórios , Recidiva , Estudos Retrospectivos , Sela Túrcica/patologia , Fatores de Tempo , Extração Dentária , Técnicas de Movimentação Dentária , Resultado do Tratamento , Adulto JovemRESUMO
PURPOSE: Clinically, we have observed the phenomenon of postoperatively accelerated orthodontic tooth movement in patients who had orthognathic surgery. This phenomenon lasts for a period of 3 to 4 months. However, the underlying mechanisms of this phenomenon have not been well studied yet. The purpose of this prospective clinical pilot study was to study the postoperative changes in bone physiology and metabolism and the corresponding responses in the dentoalveolus, such as the changes in tooth mobility. MATERIALS AND METHODS: Twenty-two consecutive adult patients who had 2-jaw orthognathic surgery were included in this study. The levels of serum alkaline phosphatase and C-terminal telopeptide of type I collagen (ICTP), as well as the tooth mobility of the maxillary and mandibular incisors based on the Periotest method (Siemens AG, Bensheim, Germany), were examined preoperatively and 1 week, 1 month, 2 months, 3 months, and 4 months postoperatively. The data were analyzed statistically. RESULTS: Both tooth mobility of the maxillary and mandibular incisors and ICTP significantly increased from 1 week to 3 months postoperatively and then decreased to their preoperative levels in the fourth month postoperatively. The changes in tooth mobility were significantly in correspondence with the changes in ICTP. The alkaline phosphatase level significantly increased from the first to fourth month postoperatively, but it was not significantly correlated to the changes in tooth mobility. CONCLUSION: The orthognathic surgery triggers a 3- to 4-month period of higher osteoclastic activities and metabolic changes in the dentoalveolus postoperatively, which possibly accelerates postoperative orthodontic tooth movement.
Assuntos
Processo Alveolar/metabolismo , Remodelação Óssea , Procedimentos Cirúrgicos Ortognáticos/métodos , Osteotomia de Le Fort , Técnicas de Movimentação Dentária/métodos , Adulto , Fosfatase Alcalina/biossíntese , Colágeno Tipo I/biossíntese , Análise do Estresse Dentário , Humanos , Incisivo/fisiopatologia , Mandíbula , Osteoclastos/fisiologia , Fragmentos de Peptídeos/biossíntese , Peptídeos , Projetos Piloto , Cuidados Pós-Operatórios , Pró-Colágeno/biossíntese , Estudos Prospectivos , Estatísticas não Paramétricas , Fatores de Tempo , Mobilidade Dentária/fisiopatologiaRESUMO
The surgery-first approach indicates that the orthognathic surgery precedes the orthodontic treatment, whereas the orthodontics-first approach indicates that the orthodontic treatment precedes the orthognathic surgery. The conventional approach is an orthodontics-first approach. The purposes of this article are to introduce the concept of the surgery-first approach and to report the guidelines for orthodontic management and model surgery without presurgical orthodontic decompensation. The surgery-first approach treats facial esthetics first and then occlusion, whereas the conventional approach treats occlusion first and then facial esthetics. The surgery-first approach uses osteotomy to solve both skeletal problems and dental compensation, and a "transitional" occlusion is set up postoperatively. Orthodontics in the surgery-first approach is a postoperatively adjunctive treatment to transfigure the transitional occlusion into the solid final occlusion. The advantages of the surgery-first approach are that 1) the patient's chief complaint, dental function, and facial esthetics are achieved and improved in the beginning of the treatment; 2) the entire treatment period is shortened to 1 to 1.5 years or fewer depending on the complexity of orthodontic treatment; and 3) the phenomenon of postoperatively accelerated orthodontic tooth movement reduces the difficulty and treatment time of orthodontic management in the surgery-first approach.
Assuntos
Má Oclusão Classe III de Angle/cirurgia , Má Oclusão Classe II de Angle/cirurgia , Ortodontia Corretiva/métodos , Procedimentos Cirúrgicos Ortognáticos/métodos , Osteotomia de Le Fort , Estética Dentária , Humanos , Má Oclusão Classe II de Angle/fisiopatologia , Má Oclusão Classe II de Angle/terapia , Má Oclusão Classe III de Angle/fisiopatologia , Má Oclusão Classe III de Angle/terapia , Modelos Anatômicos , Cuidados Pós-Operatórios , Guias de Prática Clínica como Assunto , Fatores de TempoRESUMO
To investigate the frequency of tetracycline resistance (Tet-R) lactobacilli in pig intestines, a total of 256 pig colons were analyzed and found to contain typical colonies of Tet-R lactic acid bacteria in every sample, ranging from 3.2 × 10(3) to 6.6 × 10(5) CFU/cm(2). From these samples, a total of 159 isolates of Tet-R lactobacilli were obtained and identified as belonging to 11 species, including Lactobacillus reuteri, Lactobacillus amylovorus, Lactobacillus salivarius, Lactobacillus plantarum, Lactobacillus ruminis, Lactobacillus kefiri, Lactobacillus fermentum, Lactobacillus sakei, Lactobacillus coryniformis, Lactobacillus parabuchneri and Lactobacillus letivazi. Based on the EFSA (2008) breakpoints, all isolates, after MIC analysis, were qualified as Tet-R, from which the significant high Tet-R MIC(50) and MIC(90) values indicated an ecological distribution of Tet-R lactobacilli mostly with high resistance potency in pig colons. PCR-detection identified 5 tet genes in these isolates, the most predominant one being tet (W), followed by tet (M), (L), (K), and (Q). Their detection rates were 82.0%, 22.5%, 14.4%, 8.1% and 0.9%, respectively. Noteworthily, isolates of the same species carrying identical tet gene(s) usually had a wide different MIC values. Furthermore, strain-subtyping of these isolates by REP-PCR demonstrated a notable genotypic biodiversity % (average = 62%).
Assuntos
Intestinos/microbiologia , Lactobacillus/classificação , Lactobacillus/isolamento & purificação , Resistência a Tetraciclina/genética , Animais , Antibacterianos/farmacologia , Carga Bacteriana , Proteínas de Bactérias/genética , Lactobacillus/efeitos dos fármacos , Lactobacillus/genética , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Suínos , TaiwanRESUMO
Feline mammary tumor is a relatively commonly diagnosed neoplasm in the cat. Development of new veterinary cancer therapies is limited by the shortage of in vivo models that reproduce tumor microenvironment and metastatic progression. Four feline mammary tumor orthotopic patient-derived xenograft model (PDX) successfully established in NOD-SCID gamma (NSG) mice. The overall success rate of PDX establishment was 36% (4/11). Histological, immunohistochemical, and short tandem repeat analysis showed a remarkable similarity between patient's tumor and xenograft. The tumor grafts conserve original tumor essential features, including distant metastasis. Primary FMT-1807 cell line isolated from FMT-1807PDX tumor tissue. Tumorigenicity of FMT-1807 cells expanded from PDX was assessed by orthotopic injection into NSG mice. Mice yielded tumors which preserve the lung and liver metastasis ability. This work provides a platform for FMT translational investigation.
RESUMO
Bacteroides fragilis (BF) plays a critical role in developing and maintaining the mammalian immune system. We previously found that BF colonization could prevent inflammation and tumor formation in a germ-free (GF) colitis-associated colorectal cancer (CAC) mouse model. The role of Toll-like receptor 4 (TLR4) in CAC development has not been clearly elucidated in BF mono-colonized gnotobiotic mice. The wild-type (WT) and TLR4 knockout (T4K) germ-free mice were raised with or without BF colonization for 28 days (GF/WT, GF/T4K, BF/WT, and BF/T4K) and then CAC was induced under azoxymethane (AOM)/dextran sulfate sodium (DSS) administration. The results showed that tumor formation and tumor incidence were significantly inhibited in the BF/WT group compared to those observed in the GF/WT group. However, the tumor prevention effect was not observed in the BF/T4K group unlike in the BF/WT group. Moreover, the CAC histological severity of the BF/WT group was ameliorated, but more severe lesions were found in the GF/WT, GF/T4K, and BF/T4K groups. Immunohistochemistry showed decreased cell proliferation (PCNA, ß-catenin) and inflammatory markers (iNOS) in the BF/WT group compared to those in the BF/T4K group. Taken together, BF mono-colonization of GF mice might prevent CAC via the TLR4 signal pathway.
Assuntos
Bacteroides fragilis , Neoplasias Associadas a Colite , Colite , Neoplasias Colorretais , Receptor 4 Toll-Like/genética , Animais , Azoximetano , Colite/induzido quimicamente , Colite/metabolismo , Colite/microbiologia , Colite/patologia , Neoplasias Associadas a Colite/metabolismo , Neoplasias Associadas a Colite/microbiologia , Neoplasias Associadas a Colite/patologia , Colo/metabolismo , Colo/microbiologia , Colo/patologia , Neoplasias Colorretais/etiologia , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/microbiologia , Neoplasias Colorretais/patologia , Ciclo-Oxigenase 2/metabolismo , Sulfato de Dextrana , Modelos Animais de Doenças , Vida Livre de Germes , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico Sintase Tipo II/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , beta Catenina/metabolismoRESUMO
PURPOSE: To investigate transverse dimensional changes of dental arches in surgical skeletal Class III patients with and without presurgical orthodontics. MATERIALS AND METHODS: Thirty-six patients with skeletal Class III were included and grouped into those with or without presurgical orthodontics. Eighteen patients (mean age, 22.3 +/- 3.8 years) with presurgical orthodontics (mean presurgical orthodontic treatment time, 176.3 +/- 38.3 days) were in the orthodontics-first (OF) group; the other 18 patients (mean age, 23.3 +/- 4.2 years) without presurgical orthodontics were in the surgery-first (SF) group. The posteroanterior cephalograms initially, before surgery, immediately after surgery, and 1 year after surgery were traced and analyzed. The inclination change of canines and molars was measured to interpret the changes of transverse dimension in both dental arches. Paired and unpaired t tests were performed to test intra- and intergroup differences (P < .05). RESULTS: Dental changes in transverse planes demonstrated a similar trend in both groups. The maxillary canines were buccally tilted (SF vs OF group, 1.7 degrees vs 1.9 degrees), the maxillary molars were lingually tilted (SF group vs OF group, -4.7 degrees vs -1.0 degrees), the mandibular canines were lingually tilted (SF group vs OF group, 2.9 degrees vs 2.8 degrees), and the mandibular molars were buccally tilted (SF group vs OF group, -6.1 degrees vs -5.4 degrees). CONCLUSION: The magnitude and trend of transverse dental changes in patients with surgical skeletal Class III were similar whether receiving presurgical orthodontics or not.