RESUMEN
BACKGROUND AND AIMS: Hepatic ischemia-reperfusion (HIR) injury, a common clinical complication of liver transplantation and resection, affects patient prognosis. Ring finger protein 5 (RNF5) is an E3 ubiquitin ligase that plays important roles in endoplasmic reticulum stress, unfolded protein reactions, and inflammatory responses; however, its role in HIR is unclear. APPROACH AND RESULTS: RNF5 expression was significantly down-regulated during HIR in mice and hepatocytes. Subsequently, RNF5 knockdown and overexpression of cell lines were subjected to hypoxia-reoxygenation challenge. Results showed that RNF5 knockdown significantly increased hepatocyte inflammation and apoptosis, whereas RNF5 overexpression had the opposite effect. Furthermore, hepatocyte-specific RNF5 knockout and transgenic mice were established and subjected to HIR, and RNF5 deficiency markedly aggravated liver damage and cell apoptosis and activated hepatic inflammatory responses, whereas hepatic RNF5 transgenic mice had the opposite effect compared with RNF5 knockout mice. Mechanistically, RNF5 interacted with phosphoglycerate mutase family member 5 (PGAM5) and mediated the degradation of PGAM5 through K48-linked ubiquitination, thereby inhibiting the activation of apoptosis-regulating kinase 1 (ASK1) and its downstream c-Jun N-terminal kinase (JNK)/p38. This eventually suppresses the inflammatory response and cell apoptosis in HIR. CONCLUSIONS: We revealed that RNF5 protected against HIR through its interaction with PGAM5 to inhibit the activation of ASK1 and the downstream JNK/p38 signaling cascade. Our findings indicate that the RNF5-PGAM5 axis may be a promising therapeutic target for HIR.
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Proteínas de la Membrana , Fosfoproteínas Fosfatasas , Daño por Reperfusión , Ubiquitina-Proteína Ligasas , Animales , Apoptosis , Humanos , Hígado/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Fosfoproteínas Fosfatasas/metabolismo , Daño por Reperfusión/metabolismo , Daño por Reperfusión/prevención & control , Ubiquitina-Proteína Ligasas/metabolismo , UbiquitinaciónRESUMEN
In this work, a series of novel arylamide derivatives containing piperazine moiety were designed and synthesised as tubulin polymerisation inhibitors. Among 25 target compounds, compound 16f (MY-1121) exhibited low nanomolar IC50 values ranging from 0.089 to 0.238 µM against nine human cancer cells. Its inhibitory effects on liver cancer cells were particularly evident with IC50 values of 89.42 and 91.62 nM for SMMC-7721 and HuH-7 cells, respectively. Further mechanism studies demonstrated that compound 16f (MY-1121) could bind to the colchicine binding site of ß-tubulin and directly act on ß-tubulin, thus inhibiting tubulin polymerisation. Additionally, compound 16f (MY-1121) could inhibit colony forming ability, cause morphological changes, block cell cycle arrest at the G2 phase, induce cell apoptosis, and regulate the expression of cell cycle and cell apoptosis related proteins in liver cancer cells. Overall, the promising bioactivities of compound 16f (MY-1121) make the novel arylamide derivatives have the value for further development as tubulin polymerisation inhibitors with potent anticancer activities.
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Neoplasias Hepáticas , Tubulina (Proteína) , Humanos , Apoptosis , Sitios de Unión , Piperazina , Moduladores de TubulinaRESUMEN
BACKGROUND AND AIMS: Hepatic ischemia-reperfusion (I/R) injury, which mainly involves inflammatory responses and apoptosis, is a common cause of organ dysfunction in liver transplantation (LT). As a critical mediator of inflammation and apoptosis in various cell types, the role of tripartite motif-containing (TRIM) 27 in hepatic I/R injury remains worthy of study. APPROACH AND RESULTS: This study systemically evaluated the putative role of TRIM27/transforming growth factor ß-activated kinase 1 (TAK1)/JNK (c-Jun N-terminal kinase)/p38 signaling in hepatic I/R injury. TRIM27 expression was significantly down-regulated in liver tissue from LT patients, mice subjected to hepatic I/R surgery, and hepatocytes challenged by hypoxia/reoxygenation (H/R) treatment. Subsequently, using global Trim27 knockout mice (Trim27-KO mice) and hepatocyte-specific Trim27 transgenic mice (Trim27-HTG mice), TRIM27 functions to ameliorate liver damage, reduce the inflammatory response, and prevent cell apoptosis. In parallel in vitro studies, activating TRIM27 also prevented H/R-induced hepatocyte inflammation and apoptosis. Mechanistically, TRIM27 constitutively interacted with the critical components, TAK1 and TAK1 binding protein 2/3 (TAB2/3), and promoted the degradation of TAB2/3, leading to inactivation of TAK1 and the subsequent suppression of downstream JNK/p38 signaling. CONCLUSIONS: TRIM27 is a key regulator of hepatic I/R injury by mediating the degradation of TAB2/3 and suppression of downstream TAK1-JNK/p38 signaling. TRIM27 may be a promising approach to protect the liver against I/R-mediated hepatocellular damage in transplant recipients.
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Proteínas de Unión al ADN/metabolismo , Trasplante de Hígado/efectos adversos , Hígado/irrigación sanguínea , Proteínas Nucleares/metabolismo , Daño por Reperfusión/patología , Ubiquitina-Proteína Ligasas/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Biopsia , Línea Celular , Proteínas de Unión al ADN/genética , Modelos Animales de Enfermedad , Humanos , Hígado/patología , Quinasas Quinasa Quinasa PAM/metabolismo , Masculino , Ratones , Ratones Noqueados , Proteolisis , RNA-Seq , Daño por Reperfusión/etiología , Ubiquitina-Proteína Ligasas/genéticaRESUMEN
BACKGROUND: The risk of chronic hepatitis B (CHB) infection is influenced by aberrant DNA methylation and altered nucleotide synthesis and repair, possibly caused by polymorphic variants in one-carbon metabolism genes. In the present study, we investigated the relationship between polymorphisms belonging to the one-carbon metabolic pathway and CHB infection. METHODS: A case-control study using 230 CHB patients and 234 unrelated healthy controls was carried out to assess the genetic association of 24 single nucleotide polymorphisins (SNPs) determined by mass spectrometry. RESULTS: Three SNPs, comprising rs10717122 and rs2229717 in serine hydroxymethyltransferase1/2 (SHMT2) and rs585800 in betaine-homocysteine S-methyltransferase (BHMT), were associated with the risk of CHB. Patients with DEL allele, DEL.DEL and DEL.T genotypes of rs10717122 had a 1.40-, 2.00- and 1.83-fold increased risk for CHB, respectively. Cases inheriting TA genotype of rs585800 had a 2.19-fold risk for CHB infection. The T allele of rs2229717 was less represented in the CHB cases (odds ratio = 0.66, 95% confidence interval = 0.48-0.92). The T allele of rs2229717 was less in patients with a low hepatitis B virus-DNA level compared to the control group (odds ratio = 0.49, 95% confidence interval = 0.25-0.97) and TT genotype of rs2229717 had a significant correlation with hepatitis B surface antigen level (p = 0.0195). Further gene-gene interaction analysis showed that subjects carrying the rs10717122 DEL.DEL/DEL.T and rs585800 TT/TA genotypes had a 2.74-fold increased risk of CHB. CONCLUSIONS: The results of the present study suggest that rs10717122, rs585800 and rs2229717 and gene-gene interactions of rs10717122 and rs585800 affect the outcome of CHB infection, at the same time as indicating their usefulness as a predictive and diagnostic biomarker of CHB infection.
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Betaína-Homocisteína S-Metiltransferasa/genética , Carbono/metabolismo , Glicina Hidroximetiltransferasa/genética , Hepatitis B Crónica/genética , 5-Metiltetrahidrofolato-Homocisteína S-Metiltransferasa/genética , Adenosilhomocisteinasa/genética , Adulto , Pueblo Asiatico/genética , Estudios de Casos y Controles , Metilasas de Modificación del ADN/genética , Enzimas Reparadoras del ADN/genética , Femenino , Predisposición Genética a la Enfermedad , Glicina N-Metiltransferasa/genética , Hepatitis B Crónica/metabolismo , Humanos , Masculino , Metionina Adenosiltransferasa/genética , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Proteínas Supresoras de Tumor/genéticaRESUMEN
BACKGROUND AND AIMS: Hepatic ischemia-reperfusion (I/R) injury remains a major challenge affecting the morbidity and mortality of liver transplantation. Effective strategies to improve liver function after hepatic I/R injury are limited. Six-transmembrane epithelial antigen of the prostate 3 (Steap3), a key regulator of iron uptake, was reported to be involved in immunity and apoptotic processes in various cell types. However, the role of Steap3 in hepatic I/R-induced liver damage remains largely unclear. APPROACH AND RESULTS: In the present study, we found that Steap3 expression was significantly up-regulated in liver tissue from mice subjected to hepatic I/R surgery and primary hepatocytes challenged with hypoxia/reoxygenation insult. Subsequently, global Steap3 knockout (Steap3-KO) mice, hepatocyte-specific Steap3 transgenic (Steap3-HTG) mice, and their corresponding controls were subjected to partial hepatic warm I/R injury. Hepatic histology, the inflammatory response, and apoptosis were monitored to assess liver damage. The molecular mechanisms of Steap3 function were explored in vivo and in vitro. The results demonstrated that, compared with control mice, Steap3-KO mice exhibited alleviated liver damage after hepatic I/R injury, as shown by smaller necrotic areas, lower serum transaminase levels, decreased apoptosis rates, and reduced inflammatory cell infiltration, whereas Steap3-HTG mice had the opposite phenotype. Further molecular experiments showed that Steap3 deficiency could inhibit transforming growth factor-ß-activated kinase 1 (TAK1) activation and downstream c-Jun N-terminal kinase (JNK) and p38 signaling during hepatic I/R injury. CONCLUSIONS: Steap3 is a mediator of hepatic I/R injury that functions by regulating inflammatory responses as well as apoptosis through TAK1-dependent activation of the JNK/p38 pathways. Targeting hepatocytes, Steap3 may be a promising approach to protect the liver against I/R injury.
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Proteínas de Ciclo Celular/fisiología , Hepatocitos/enzimología , Hígado/irrigación sanguínea , Quinasas Quinasa Quinasa PAM/antagonistas & inhibidores , Oxidorreductasas/fisiología , Daño por Reperfusión/prevención & control , Animales , Apoptosis , Proteínas de Ciclo Celular/deficiencia , Inflamación/etiología , Proteínas Quinasas JNK Activadas por Mitógenos/fisiología , Quinasas Quinasa Quinasa PAM/fisiología , Masculino , Ratones , Oxidorreductasas/deficiencia , Daño por Reperfusión/patología , Transducción de Señal , Proteínas Quinasas p38 Activadas por Mitógenos/fisiologíaRESUMEN
OBJECTIVE: To develop an indirect immunofluorescence assay (IFA) kit for detecting anti-Toxoplasma gondii IgG. METHODS: Based on the established IFA method, we established an IFA kit for the detection of human T. gondii infection. The optimal working concentrations of T. gondii IgG-positive human serum and FITC-labeled goat anti-human IgG antibody were determined. Sensitivity, specificity, reproducibility, and storage period of this kit were studied, and compared with an imported kit. RESULTS: The optimal working concentration of T. gondii IgG-positive human serum and FITC-labeled goat anti-human IgG antibody was 1:40 and 1:100, respectively. The maximum dilution of T. gondii IgG-positive human serum that the kit can detect was 1:640. No cross reaction was observed with sera from patients with vivax malaria, falciparum malaria, schistosomiasis, echinococcosis, or cysticercosis. Cross reaction was observed to the rheumatoid factor positive sera. The sensitivity, specificity, positive predictive value, negative predictive value, concordance, Youden index of this kit was 90.9%, 100%, 100%, 96.2%, 97.2%, and 0.91, respectively; and that of the imported kit was 100%, 98%, 95.7%, 100%, 98.6%, and 0.98, respectively. There was no significant difference in sensitivity and specificity between the two kits (P > 0.05). CONCLUSION: The IFA kit shows adequate sensitivity and specificity for detection of anti-T. gondii IgG.
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Anticuerpos Antiprotozoarios/sangre , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Inmunoglobulina G/sangre , Toxoplasmosis/diagnóstico , Humanos , Juego de Reactivos para Diagnóstico , Sensibilidad y Especificidad , Toxoplasma/inmunología , Toxoplasmosis/sangreRESUMEN
Normothermic machine perfusion (NMP) could provide a curative treatment to reduce biliary injury in donation after cardiac death (DCD) donor livers; however, the underlying mechanisms remain poorly understood. In a rat model, our study compared air-oxygenated NMP to hyperoxygenated NMP and found that air-oxygenated NMP improved DCD functional recovery. Here, we found that the charged multivesicular body protein 2B (CHMP2B) expression was substantially elevated in the intrahepatic biliary duct endothelium of the cold-preserved rat DCD liver after air-oxygenated NMP or in biliary endothelial cells under hypoxia/physoxia. CHMP2B knockout (CHMP2B-/-) rat livers showed increased biliary injury after air-oxygenated NMP, indicated by decreased bile production and bilirubin level, elevated biliary levels of lactate dehydrogenase and gamma-glutamyl transferase. Mechanically, we demonstrated that CHMP2B was transcriptionally regulated by Kruppel-like transcription factor 6 (KLF6) and alleviated biliary injury through decreasing autophagy. Collectively, our results suggested that air-oxygenated NMP regulates CHMP2B expression through the KLF6, which reduces biliary injury by inhibiting autophagy. Targeting the KLF6-CHMP2B autophagy axis may provide a solution to reducing biliary injury in DCD livers undergoing NMP.
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Células Endoteliales , Trasplante de Hígado , Ratas , Animales , Cuerpos Multivesiculares , Trasplante de Hígado/métodos , Preservación de Órganos/métodos , Hígado , Perfusión/métodos , MuerteRESUMEN
OBJECTIVE: To explore the associations between the single nucleotide polymorphism of human mismatch repair gene hMLH1 and the papillary thyroid carcinoma (PTC) in Chinese Han people. METHODS: A hospital based 1:1 matched case-control study was carried out. The single nucleotide polymorphism (-93G > A, 1151T > A and 655A > G) for 204 pairs of cases with PTC as well as healthy controls was identified by PCR-RFLP, PCR-ASO and DNA sequencing. RESULTS: With univariate analysis, we found that compared to 1151TT genotype, the TA genotype could increase the PTC risk marginally, with odds ratio (OR) of 2.15 (95%CI: 0.99 - 4.85); While the mutant genotype TA + AA could increase the PTC risk statistically significant, with OR of 2.15(95%CI: 1.02 - 4.69). With 2 x 4 cross-over study, we found that compared to -93GG and 1151TT genotypes, individuals with both -93GA + AA and 1151TA + AA could increase the PTC risk marginally, with OR of 2.50 (95%CI: 0.96 - 6.67); While, compared to 655AA and 1151TT genotypes, individuals with both 655AA and 1151TA + AA could increase the PTC risk statistically significant, with OR of 2.50 (95%CI: 1.02 - 4.73). Multivariate and conditional logistic regression analysis showed the genotype of 1151TA, the history of receiving CT diagnosis, the history of tumor, the negative life events and eating seafood frequently could increase the risk of PTC, with OR of 6.79 (95%CI: 3.18 - 14.49), 3.35 (95%CI: 1.93 - 5.80), 39.03 (95%CI: 3.70 - 41.60) and 3.98 (95%CI: 1.81 - 8.73); While, eating fruit frequently could decrease the PTC risk. CONCLUSION: The 1151TA + AA genotype, the history of receiving CT diagnosis, the history of tumor, the negative life events and eating seafood frequently were the risk factors of PTC, while eating fruit frequently was the protective factor.
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Proteínas Adaptadoras Transductoras de Señales/genética , Carcinoma Papilar/genética , Predisposición Genética a la Enfermedad , Proteínas Nucleares/genética , Polimorfismo Genético , Neoplasias de la Tiroides/genética , Adulto , Pueblo Asiatico/genética , Carcinoma Papilar/epidemiología , Carcinoma Papilar/etnología , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Masculino , Análisis por Apareamiento , Persona de Mediana Edad , Homólogo 1 de la Proteína MutL , Neoplasias de la Tiroides/epidemiología , Neoplasias de la Tiroides/etnologíaRESUMEN
Hepatocellular carcinoma (HCC), a type of malignant liver tumor, has a grim prognosis. As a functional protein, synaptopodin-2 (SYNPO2) has been associated with malignancy; however, the expression profile and function of SYNPO2 in HCC remains unknown. Herein, we revealed that SYNPO2 was transcriptionally downregulated in HCC tissues from both The Cancer Genome Atlas cohort and our cohort, and was also decreased at the translational level as determined by western blotting and immunohistochemical staining. Furthermore, reduced SYNPO2 expression correlated significantly with short overall survival and recurrence free survival of HCC patients. Restoring SYNPO2 expression inhibited the proliferation and aggressiveness of hepatocarcinoma cells. Mechanistically, increasing the ratio of cytoplasmic SYNPO2 to nuclear SYNPO2 was positively associated with recurrence rate in HCC patients; calcineurin (CaN) activity positively correlated with cytoplasmic SYNPO2 levels in HCC tissues; and nuclear-cytoplasmic translocation of SYNPO2 was induced by CaN to facilitate metastasis of HCC through assembly of peripheral actin bundles. In short, our findings uncover a novel role of SYNPO2 in HCC metastasis via the CaN/SYNPO2/F-actin axis, and indicate that SYNPO2 may serve as a possible prognostic marker and novel therapeutic target.
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Calcineurina/metabolismo , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/patología , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Animales , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Línea Celular Tumoral , Movimiento Celular , Núcleo Celular/metabolismo , Proliferación Celular , Citoplasma/metabolismo , Regulación hacia Abajo , Femenino , Regulación Neoplásica de la Expresión Génica , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Masculino , Ratones , Metástasis de la Neoplasia , Trasplante de Neoplasias , Pronóstico , Transporte de Proteínas , Análisis de SupervivenciaRESUMEN
Thirty Kunming mice were randomly divided into three groups named as dihydroartemisinin roup (A), dihydroartemisinin and azithromycin group (B), and control group (C). Each mouse was infected intraperitoneally with 2x10(3) Toxoplasma gondii tachyzoites. Eight hours after infection, the mice of groups A and B were treated twice a day for 4 days with 75 mg/kg of dihydroartemisinin. At 24 hours post infection, those in group B were treated once daily for 4 days with 200 mg/kg of azithromycin. At 96 hours post infection, ascites was taken from one mouse each group and the tachyzoites were collected. The ultrastructure of tachyzoites was observed by conventional transmission electron microscopy. It was found that the tachyzoites in groups A and B showed edema and enlarged, the cell membrane became indistinct, broken or damaged; fat droplets in the cytoplasm increased, and vacuoles were formed. Similar changes were not seen in the control group.
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Artemisininas/farmacología , Azitromicina/farmacología , Toxoplasma/efectos de los fármacos , Toxoplasma/ultraestructura , Toxoplasmosis Animal/parasitología , Animales , Antiprotozoarios/farmacología , Femenino , Ratones , Ratones EndogámicosRESUMEN
OBJECTIVE: To explore the relationship of the genetic polymorphisms and the haplotypes in hMLH1 and hMSH3 gene with the risk of papillary thyroid carcinoma (PTC) in Chinese Hans. METHODS: A hospital based 1:1 matched case-control study was carried out. The polymorphisms for 204 pairs of PTC cases and healthy controls were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allele specific oligonucleotide (PCR-ASO) assays. RESULTS: (1) The PTC risk was marginally increased in the hMLH1 1151TA genotype, with odds ratio (OR) of 2.15 (95%CI: 0.99-4.85); the PTC risk was significantly increased in the mutant genotype 1151TA+AA, with OR of 2.15 (95%CI: 1.02-4.69); (2) The haplotypes of -93G, 1151A, 655A in the hMLH1 gene could increase the PTC risk, with OR of 2.67 (95%CI: 1.16-6.53, P=0.011), compared with the haplotype of -93G, 1151T, 655A; (3) Compared to 3124A, 2835G haplotype in hMSH3 gene, the 3124G, 2835A haplotype could increase the PTC risk marginally, with OR of 3.08 (95%CI: 0.92-13.25). CONCLUSION: The 1151T/A polymorphism in hMLH1 was associated with PTC; both the haplotype of -93G, 1151A, 655A in hMLH1 and the 3124G, 2835A haplotype in hMSH3 were associated with PTC.
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Adenocarcinoma Papilar/genética , Haplotipos/genética , Polimorfismo de Nucleótido Simple , Adulto , Femenino , Predisposición Genética a la Enfermedad , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Riesgo , Neoplasias de la Tiroides/genéticaRESUMEN
Mice were divided into 3 groups: heavy infection group with 80 mice each was fed with 400 muscle larvae of Trichinella spiralis, light infection group with 60 mice each was fed by 200 larvae, and uninfected control (60 mice) . The content of Cu, Zn and Fe in the dorsal hair samples was measured in the week of 1, 3, 5, 7, 9, 11, 13 and 15 after infection. Results indicated that the content of Zn, Cu and Fe in the two experimental groups reduced considerably in comparison to the control (P < 0.05), especially for that of Zn and Cu. Lower content was found in the heavily infected mice than in those with light infection (P < 0.05).
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Cabello/química , Oligoelementos/análisis , Trichinella spiralis/fisiología , Triquinelosis/metabolismo , Animales , Cobre/análisis , Cabello/parasitología , Interacciones Huésped-Parásitos , Hierro/análisis , Ratones , Ratones Endogámicos , Triquinelosis/parasitología , Zinc/análisisRESUMEN
OBJECTIVE: To study the change of enzymes and effect of garlicin treatment on the change in bronchoalveolar lavage fluid (BALF) of rats with Pneumocystis carinii pneumonia (PCP). METHODS: Wistar rats were injected intramuscularly continually with dexamethasone to establish the rat model of PCP. The experimental rats (group A) were injected intramuscularly with garlicin at a dose of 10 mg/(kg x d) for 5 days in the 3rd, 6th and 9th week respectively, and SMZ/TMP therapy group (B), PCP infected group (C) and normal group (D) were established as controls. Three days after the last treatment, the rats of all groups were killed and BALF was collected without contamination and enzymes AST, ALF, CHE, ALP, LDH, CK, CKMB, HBDH, AFU, 5'NT, ADA were examined. RESULTS: The ALP level in group C [(573.41 +/- 350.63)U/L] was significantly higher than that in group D [(210.56 +/- 114.41) U/L] (q = 4.682, P < 0.01), group A [(392.07 +/- 217.57) U/L] (q = 3.851, P < 0.05), and group B [(325.21 +/- 180.65) U/L] (q = 4.380, P < 0.01); the level of CK, CKMB and 5'NT in group C [948.94 +/- 403.43, 489.47 +/- 254.46 and (6.76 +/- 3.11) U/L respectively] was higher than those in group D [426.22 +/- 319.00, 213.33 +/- 144.54 and (3.22 +/- 1.20) U/L] (q = 4.696, 3.784, 3.812, P< 0.05); there was no significant difference in the level of AST, ALT, CHE, LDH, HBDH, AFU and ADA among the four groups (F = 1.852, 0.958, 2.470, 1.423, 1.178, 1.342, 0.611, P > 0.05). CONCLUSIONS: The level of ALP, CK, CKMB but the ALP level decreases distinctly after the garlicin and 5'NT increases evidently in BALF of PCP infected rats, but the ALP level decreases distinctly after the garlicin treatment.
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Compuestos Alílicos/farmacología , Disulfuros/farmacología , Infecciones por Pneumocystis/tratamiento farmacológico , Neumonía por Pneumocystis/tratamiento farmacológico , Alanina Transaminasa/metabolismo , Compuestos Alílicos/uso terapéutico , Animales , Aspartato Aminotransferasas/metabolismo , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/microbiología , Dexametasona , Modelos Animales de Enfermedad , Disulfuros/uso terapéutico , Femenino , Humanos , L-Lactato Deshidrogenasa/metabolismo , Infecciones por Pneumocystis/inducido químicamente , Infecciones por Pneumocystis/metabolismo , Pneumocystis carinii/efectos de los fármacos , Neumonía por Pneumocystis/inducido químicamente , Neumonía por Pneumocystis/metabolismo , Ratas , Ratas WistarRESUMEN
Bromodomain and Extra-Terminal Domain (BET) inhibitors, such as JQ1 have emerged as novel drug candidates and are being enthusiastically pursued in clinical trials for the treatment of cancer. However, many solid cancers are resistance to BET inhibitors. To explore methods for improving the therapeutic potential of BET inhibitors, we investigated the combinational activity of JQ1 with Oridonin, a bioactive molecules derived from Traditional Chinese Medicine in hepatocellular carcinoma (HCC) cells. Our results showed that Oridonin synergistically enhanced the abilities of JQ1 to inhibit cell viability in HCC cells and, significantly augmented JQ1-triggered apoptosis in HCC cells and in HCC cancer stem-like cells. Moreover, Oridonin dose-dependently inhibited the expression of several anti-apoptotic proteins, such as Bcl-2, Mcl-1, and x-linked inhibitor of apoptosis (xIAP) in HCC cells. Cell fractionation and western blotting analysis showed that the enhancement of apoptosis by Oridonin was associated with cytochrome c release, activation of caspase-9, -3 and cleavage of PARP, indicating the activation of mitochondrial apoptosis pathway. Altogether, our findings demonstrate that Oridonin may be used to effectively enhance the sensitivity of BET inhibitors in HCC therapy via downregulation of the expression of multiple anti-apoptotic proteins.
RESUMEN
Bromodomain 4 (BRD4) is an epigenetic regulator that, when inhibited, has anti-cancer effects. In this study, we investigated whether BRD4 could be a target for treatment of human hepatocellular carcinoma (HCC). We show that BRD4 is over-expressed in HCC tissues. Suppression of BRD4, either by siRNA or using JQ1, a pharmaceutical BRD4 inhibitor, reduced cell growth and induced apoptosis in HCC cell lines while also slowing HCC xenograft tumor growth in mice. JQ1 treatment induced G1 cell cycle arrest by repressing MYC expression, which led to the up-regulation of CDKN1B (P27). JQ1 also de-repressed expression of the pro-apoptotic BCL2L11 (BIM). Moreover, siRNA knockdown of BIM attenuated JQ1-triggered apoptosis in HCC cells, suggesting an essential role for BIM in mediating JQ1 anti-HCC activity.
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Azepinas/farmacología , Proteína 11 Similar a Bcl2/metabolismo , Carcinoma Hepatocelular/prevención & control , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Neoplasias Hepáticas/prevención & control , Proteínas Nucleares/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-myc/metabolismo , Factores de Transcripción/antagonistas & inhibidores , Triazoles/farmacología , Animales , Apoptosis/efectos de los fármacos , Proteína 11 Similar a Bcl2/genética , Western Blotting , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Proteínas de Ciclo Celular , Proliferación Celular/efectos de los fármacos , Inmunoprecipitación de Cromatina , Citometría de Flujo , Humanos , Técnicas para Inmunoenzimas , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Ratones , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The aim was to investigate the effects of the GABAB receptor antagonist, CGP46381, on form-deprivation myopia (FDM) in guinea pigs. Twenty-four guinea pigs had monocular visual deprivation induced using a diffuser for 11 days (day 14 to 25). The deprived eyes were treated with daily subconjunctival injections (100 µl) of either 2% CGP46381, 0.2% CGP46381, or saline or received no injection. The fellow eyes were left untreated. Another six animals received no treatment. At the start and end of the treatment period, ocular refractions were measured using retinoscopy and vitreous chamber depth (VCD) and axial length (AL) using A-scan ultrasound. All of the deprived eyes developed relative myopia (treated versus untreated eyes, P < 0.05). The amount of myopia was significantly affected by the drug treatment (one-way ANOVA, P < 0.0001). The highest dose tested, 2% CGP46381, significantly inhibited myopia development compared to saline (2% CGP46381: -1.08 ± 0.40 D, saline: -4.33 ± 0.67 D, P < 0.01). The majority of these effects were due to less AL (2% CGP46381: 0.03 ± 0.01 mm, saline: 0.13 ± 0.02 mm, P < 0.01) and VCD (2% CGP46381: 0.02 ± 0.01 mm, saline: 0.08 ± 0.01 mm, P < 0.01) elongation. The lower dose tested, 0.2% CGP46381, did not significantly inhibit FDM (P > 0.05). Subconjunctival injections of CGP46381 inhibit FDM development in guinea pigs in a dose-dependent manner.
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Percepción de Forma/efectos de los fármacos , Antagonistas de Receptores de GABA-B/farmacología , Antagonistas de Receptores de GABA-B/uso terapéutico , Miopía/tratamiento farmacológico , Miopía/fisiopatología , Ácidos Fosfínicos/uso terapéutico , Receptores de GABA-B/metabolismo , Animales , Longitud Axial del Ojo/efectos de los fármacos , Longitud Axial del Ojo/fisiopatología , Cobayas , Ácidos Fosfínicos/farmacología , Refracción Ocular/efectos de los fármacos , Cuerpo Vítreo/efectos de los fármacos , Cuerpo Vítreo/fisiopatologíaRESUMEN
OBJECTIVE: To explore the single nucleotide polymorphisms (SNPs) and point mutations of ataxia telangiectasia mutated (ATM) gene in Chinese of Han ethnicity. METHODS: The target fragments of the exon 39, exon 61 and exon 63 of ATM gene were first amplified using PCR technique, then screened for the SNPs and point mutations using single strand conformation polymorphism (SSCP) technique, finally the representative bands were sequenced for the verification of the new single nucleotide polymorphisms and point mutations using the automatic DNA sequencing technique. RESULTS: Six new SNPs were found in the exon 39, intron 61 and intron 63 of ATM gene. They are respectively the A/T polymorphisms at nucleotide 5689 and 5691 in exon 39, the T/G polymorphisms at nucleotide +69 and +99, the A/G polymorphism at nucleotide +94 in intron 61, the G/C polymorphism at nucleotide +17 in intron 63. Five new point mutations were found in the exon 61, intron 62 and exon 63 of ATM gene. They are respectively the T/G transversion at nucleotide 8618 in exon 61, the T/G transversion at nucleotide -13 in intron 62, the T/G transversion at nucleotide 8793, the G/A transitions at nucleotide 8816 and 8848 in exon 63. The known three SNPs, the G/A polymorphism at nucleotide 5557 in exon 39, the T/C polymorphism at nucleotide +104 in intron 61 and the T/C polymorphism at nucleotide -55 in intron 62, were verified in Chinese of Han ethnicity. CONCLUSION: There are great differences in the SNPs of ATM gene between Chinese of Han ethnicity and Caucasian.
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Pueblo Asiatico/genética , Ataxia Telangiectasia/genética , Proteínas de Ciclo Celular/genética , Proteínas de Unión al ADN/genética , Mutación Puntual , Polimorfismo de Nucleótido Simple/genética , Proteínas Serina-Treonina Quinasas/genética , Proteínas Supresoras de Tumor/genética , Proteínas de la Ataxia Telangiectasia Mutada , China/etnología , Exones/genética , Humanos , Intrones/genética , Polimorfismo Conformacional Retorcido-Simple , Análisis de Secuencia de ADN , Población BlancaRESUMEN
OBJECTIVE: To assess the therapeutic effect of garlicin on Pneumocystis carinii pneumonia (PCP) of immunosuppressed rats. METHODS: The Wistar rats were injected intramuscularly continually with dexamethasone for eight weeks to establish the rat model of PCP. The rats were treated with garlicin, meanwhile control groups without treatment and with SMZ-TMP treatment were established respectively in PCP model. The efficacy was evaluated based on the lung weight, lung/body weight ratio and mean cyst number per 100 fields in lung print smear. RESULTS: The mean lung weight of the rats in garlicin is 1.73 +/- 0.17, lung/body weight ratio is 0.84 +/- 0.12, they are obvious lower than 2.31 +/- 0.35 and 1.25 +/- 0.35 of control (P <0.01). The numbers of cysts in lung print smear is reduced 62.9% compared with control. The results is similar to that with SMZ-TMP. CONCLUSION: Garlicin shows an obvious therapeutic effect on Pneumocystis carinii pneumonia in rats with an efficacy similar to that with SMZ-TMP.
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Compuestos Alílicos/uso terapéutico , Antifúngicos/uso terapéutico , Disulfuros/uso terapéutico , Infecciones por Pneumocystis/tratamiento farmacológico , Animales , Dexametasona , Femenino , Ratas , Ratas WistarRESUMEN
OBJECTIVE: To study the antiproliferation effect on HepG2 cells and its underlying mechanism of the active chemical composition of the Viburnum Odoratissimum. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and trypan blue dye exclusion assay were used to assess the effect of vibsane-type diterpenoids on the proliferation of various tumor cells. Alterations in cell cycle and apoptosis were determined by flowcytometry. The enzymatic activity of caspase-3/7 was measured by Apo-ONE homogeneous Caspase-3/7 Assay kit. RESULTS: Compound 1 #, a vibsane-type diterpenoid, was found to significantly inhibit the growth of HepG2 cells by anticancer proliferation activity screening. It was demonstrated that the modified groups on side chain coupled to C11 site affected the cell growth-inhibition activity of compounds by structure-activity analysis. In addition, HepG2 cell line was most sensitive to compound 1 #, which induced growth arrest of HepG2 cells in a dose- and time-dependent manner. Study on the mechanisms underlying these effects indicated that compound 1 # induced significant G0/G1 phase arrest of HepG2 cells in a time- and concentration-dependent manner. Meanwhile, It was found that higher concentrations of compound (5-10 micromol/L) caused evident increase in the unmber of apoptotic cells and dose-dependent activation of caspase-3/7. CONCLUSION: Vibsane-type diterpenoids could significantly inhibit the growth of HCC HepG2 cells. Induction of cell cycle arrest and apoptosis may play important roles in their anticancer effects.
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Proliferación Celular/efectos de los fármacos , Diterpenos/farmacología , Viburnum/química , Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Células Hep G2 , HumanosRESUMEN
OBJECTIVE: To observe the changes of alveolar macrophage (AM) phagocytosis and the levels of TNF-alpha and IL-6 in AM supernatants and bronchoalveolar lavage fluid (BALF) of rats with Pneumocystis pneumonia (PCP). METHODS: Wistar rats were injected with dexamethasone intramuscularly and continually to establish the model of PCP, the AM phagocytosis and levels of TNF-alpha and IL-6 in AM supernatants and BALF of rats with PCP were detected, meanwhile, the normal controls were set. RESULTS: The phagocyting percentage [(20.61 +/- 2.04)%] and phagocyting index (0.25 +/- 0.21) of the PCP group were significantly lower than those [(25.45 +/- 3.1)% and (0.31 +/- 0.16)] of the control group (P < 0.05), TNF-alpha (16.84 +/- 0.86) pg/ml and IL-6 (1.05 +/- 0.19) pg/ml in BALF of the PCP group were significantly higher than those [(12.48 +/- 0.84) pg/ml and (0.86 +/- 0.11) pg/ml] of the control group (P < 0.05), but there were no significant differences for the levels of TNF-alpha and IL-6 in AM supernatants between the two groups (P > 0.05). CONCLUSION: AM phagocytosis reduces evidently of PCP rats, but the levels of TNF-alpha and IL-6 in BALF rise evidently.