RESUMEN
Novel biomarkers for pancreatic adenocarcinoma are urgently needed because of its poor prognosis. Here, by using The Cancer Genome Atlas (TCGA) RNA-seq data, we evaluated the prognostic values of the differentially expressed miRNAs and constructed a five-miRNA signature that could effectively predict patient overall survival (OS). The Kaplan-Meier overall survival curves of two groups based on the five miRNAs were notably different, showing overall survival in 10.2% and 47.8% at five years for patients in high-risk and low-risk groups, respectively. The ROC curve analysis achieved AUC of 0.775, showing good sensitivity and specificity of the five-miRNA signature model in predicting pancreatic adenocarcinoma patient survival risk. The functional enrichment analysis suggested that the target genes of the miRNA signature may be involved in various pathways related to cancer, including PI3K-Akt, TGF-ß, and pluripotent stem cell signaling pathways. Finally, we analyzed expression of the five specific miRNAs in the miRNA signature, and validated the reliability of the results in 20 newly diagnosed pancreatic adenocarcinoma patients using qRT-PCR. The expression results of qRT-PCR were consistent with the TCGA results. Taken together, these findings suggested that the five-miRNA signature (hsa-miR-203, hsa-miR-424, hsa-miR-1266 hsa-miR-1293, and hsa-miR-4772) could be used as a prognostic marker for pancreatic adenocarcinoma.
Asunto(s)
Adenocarcinoma/mortalidad , Biomarcadores de Tumor/genética , Bases de Datos Factuales , MicroARNs/genética , Neoplasias Pancreáticas/mortalidad , Adenocarcinoma/genética , Adenocarcinoma/patología , Perfilación de la Expresión Génica , Humanos , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , Pronóstico , Curva ROC , Tasa de SupervivenciaRESUMEN
Cholangiocarcinoma (CCA) is a cancer type with high postoperative relapse rates and poor long-term survival largely due to tumor invasion, distant metastasis, and multidrug resistance. Deregulated microRNAs (miRNAs) are implicated in several cancer types including CCA. The specific roles of the miRNA let-7c in cholangiocarcinoma are not known and need to be further elucidated. In our translational study we show that microRNA let-7c expression was significantly downregulated in human cholangiocarcinoma tissues when compared to adjacent tissues of the same patient. Let-7c inhibited the tumorigenic properties of cholangiocarcinoma cells including their self-renewal capacity and sphere formation in vitro and subcutaneous cancer cell growth in vivo. Ectopic let-7c overexpression suppressed migration and invasion capacities of cholangiocarcinoma cell lines in vitro, however, promoted distant invasiveness in vivo. Furthermore, we found that let-7c regulated the aforementioned malignant biological properties, at least in part, through regulation of EZH2 protein expression and through the DVL3/ß-catenin axis. The miRNA let-7c thus plays an important dual role in regulating tumorigenic and metastatic abilities of human cholangiocarcinoma through mechanisms involving EZH2 protein and the DVL3/ß-catenin axis.
Asunto(s)
Neoplasias de los Conductos Biliares/genética , Carcinogénesis/genética , Colangiocarcinoma/genética , Proteína Potenciadora del Homólogo Zeste 2/genética , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Animales , Neoplasias de los Conductos Biliares/metabolismo , Neoplasias de los Conductos Biliares/patología , Carcinogénesis/metabolismo , Carcinogénesis/patología , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Colangiocarcinoma/metabolismo , Colangiocarcinoma/patología , Proteínas Dishevelled/genética , Proteínas Dishevelled/metabolismo , Proteína Potenciadora del Homólogo Zeste 2/antagonistas & inhibidores , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , MicroARNs/metabolismo , Invasividad Neoplásica , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Transducción de Señal , Carga Tumoral , Ensayos Antitumor por Modelo de Xenoinjerto , beta Catenina/genética , beta Catenina/metabolismoRESUMEN
AIM: To examine the role of microRNA 1181 (miR-1181) in invasion and proliferation in pancreatic cancer. METHODS: We analyzed the expression of miR-1181 in several pancreatic cancer cell lines and generated stable MIA-PaCa-2 and PANC-1 cell lines with up-regulated miR-1181 expression using an adenovirus delivery system. We then investigated miR-1181's effect on invasion and proliferation of pancreatic cancer cells by transwell assay, wound healing assay, cell counting kit-8 assay and colony-forming assay, and explored any underlying mechanisms by western bolt. Beyond that, we observed the change of the PANC-1 cell's cytoskeleton by immunofluorescence staining. RESULTS: Our data showed that miR-1181 was relatively down-regulated in pancreatic cancer cell lines compared with normal pancreatic ductal epithelial cells. And miR-1181 inhibited the migration, invasion and proliferation activities of MIA-PaCa-2 and PANC-1 cells. Notably, after over-expressing of miR-1181 in PANC-1 cells, F-actin depolymerized. Immunofluorescence staining shows decreased F-actin and ß-tubulin expression in PANC-1 cells over-expressing miR-1181 compared with the control cells. Furthermore, we found that over-expressing miR-1181 inhibited the expression of signal transducer and activator of transcription 3 (STAT3) while knocking-down miR-1181 up-regulated the expression of STAT3. Knocking-down miR-1181 promoted the invasion and proliferation of pancreatic cancer cells. And inhibition of STAT3 blocked the promotion effects of knocking-down miR-1181 on proliferation and invasion in pancreatic cancer. CONCLUSION: Together our findings suggest that miR-1181 may be involved in pancreatic cancer cell invasion and proliferation by targeting STAT3 and indicate that miR-1181 may be a potential therapeutic agent for pancreatic cancer.
Asunto(s)
Regulación Neoplásica de la Expresión Génica , MicroARNs/metabolismo , Neoplasias Pancreáticas/metabolismo , Factor de Transcripción STAT3/metabolismo , Adenoviridae , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Regulación hacia Abajo , Humanos , Invasividad Neoplásica , TransfecciónRESUMEN
Pancreatic cancer is the fourth most common cause of cancer mortality worldwide. Furthermore, patients with pancreatic cancer experience limited benefit from current chemotherapeutic approaches because of drug resistance. Therefore, an effective therapeutic strategy for patients with pancreatic cancer is urgently required. Deguelin is a natural chemopreventive drug that exerts potent antiproliferative activity in solid tumors by inducing cell death. However, the molecular mechanisms underlying this activity have not been fully elucidated. Here we show that deguelin blocks autophagy and induces apoptosis in pancreatic cancer cells in vitro. Autophagy induced by doxorubicin plays a protective role in pancreatic cancer cells, and suppressing autophagy by chloroquine or silencing autophagy protein 5 enhanced doxorubicin-induced cell death. Similarly, inhibition of autophagy by deguelin also chemosensitized pancreatic cancer cell lines to doxorubicin. These findings suggest that deguelin has potent anticancer effects against pancreatic cancer and potentiates the anti-cancer effects of doxorubicin. These findings provide evidence that combined treatment with deguelin and doxorubicin represents an effective strategy for treating pancreatic cancer.
Asunto(s)
Anticarcinógenos/farmacología , Autofagia/efectos de los fármacos , Doxorrubicina/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Rotenona/análogos & derivados , Apoptosis/efectos de los fármacos , Biomarcadores , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Humanos , Neoplasias Pancreáticas/metabolismo , Rotenona/farmacologíaRESUMEN
BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are heterogeneous cell types that suppress T-cell responses in cancer patients and animal models, some MDSC subpopulations are increased in patients with pancreatic cancer. The present study was to investigate a specific subset of MDSCs in patients with pancreatic cancer and the mechanism of MDSCs increase in these patients. METHODS: Myeloid cells from whole blood were collected from 37 patients with pancreatic cancer, 17 with cholangiocarcinoma, and 47 healthy controls. Four pancreatic cancer cell lines were co-cultured with normal peripheral blood mononuclear cells (PBMCs) to test the effect of tumor cells on the conversion of PBMCs to MDSCs. Levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and arginase activity in the plasma of cancer patients were analyzed by enzyme-linked immunosorbent assay. RESULTS: CD14+/CD11b+/HLA-DR- MDSCs were increased in patients with pancreatic or bile duct cancer compared with those in healthy controls, and this increase was correlated with clinical cancer stage. Pancreatic cancer cell lines induced PBMCs to MDSCs in a dose-dependent manner. GM-CSF and arginase activity levels were significantly increased in the serum of patients with pancreatic cancer. CONCLUSIONS: MDSCs were tumor related: tumor cells induced PBMCs to MDSCs in a dose-dependent manner and circulating CD14+/CD11b+/HLA-DR- MDSCs in pancreatic cancer patients were positively correlated with tumor burden. MDSCs might be useful markers for pancreatic cancer detection and progression.
Asunto(s)
Neoplasias de los Conductos Biliares/inmunología , Colangiocarcinoma/inmunología , Leucocitos Mononucleares/inmunología , Células Mieloides/inmunología , Neoplasias Pancreáticas/inmunología , Escape del Tumor , Anciano , Arginasa/sangre , Neoplasias de los Conductos Biliares/sangre , Neoplasias de los Conductos Biliares/patología , Biomarcadores de Tumor/sangre , Antígeno CD11b/sangre , Estudios de Casos y Controles , Línea Celular Tumoral , Colangiocarcinoma/sangre , Colangiocarcinoma/patología , Técnicas de Cocultivo , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/sangre , Antígenos HLA-DR/sangre , Humanos , Leucocitos Mononucleares/metabolismo , Receptores de Lipopolisacáridos/sangre , Masculino , Persona de Mediana Edad , Células Mieloides/metabolismo , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/patología , Carga TumoralRESUMEN
The expression levels of hypoxia-inducible factor 1alpha (HIF-1α) and HIF-2α in pancreatic cancer (PC) and their association with clinicopathologic characteristics were investigated in order to elucidate their roles in the development of PC. HIF-1α and HIF-2α mRNA levels in 20 patients with PC were detected by quantitative real-time polymerase chain reaction. The expression of HIF-1α and HIF-2α protein in samples from other 90 patients with PC was measured by immunohistochemistry. Correlations between the expression of HIF-1α or HIF-2α and clinicopathologica features and prognosis were analyzed. The expression of both HIF-1α and HIF-2α mRNA was up-regulated in most cancer tissues (P<0.05). HIF-1α staining was weakly positive in most cancer tissues and strongly positive in adjacent pancreas tissues (P<0.05). Clinicopathologic analysis revealed that relatively strong HIF-1α expression in cancer tissues was related to greater invasion (P<0.05), higher tumor pathologic stage (P<0.05), higher American Joint Committee on Cancer (AJCC) stage (P<0.05) and shorter overall survival time (P<0.05). Conversely, HIF-2α staining was strongly positive in most cancer tissues and weakly positive in adjacent pancreas tissues. Clinicopathologic analysis revealed that relatively strong HIF-2α expression in cancer tissues was related to less invasion (P<0.05), lower tumor pathologic stage (P<0.05), lower AJCC stage (P<0.05) and longer overall survival time (P<0.05). Moreover, the HIF-1α(high)/HIF-2α(low) group showed a shorter survival time than the HIF-1α(low)/HIF-2α(high) group. In conclusion, although HIF-1α and HIF-2α mRNA expression patterns are the same, their protein expression patterns are significantly different and they play different roles in PC. Combined analysis of HIF-1α and HIF-2α expression might be useful to predict the prognosis of patients with PC.
Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Neoplasias Pancreáticas/metabolismo , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Neoplasias Pancreáticas/patología , Pronóstico , ARN Mensajero/genéticaRESUMEN
Inositol phosphorylceramide (IPC) synthase is the key enzyme with highly conserved sequences, which is involved in fungal sphingolipid biosynthesis. The antibiotic aureobasidin A (AbA) induces the death of fungi through inhibiting IPC synthase activity. The mutations of AUR1 gene coding IPC synthase in fungi and protozoa causes a resistance to AbA. However, the mechanism of AbA resistance is still elusive. In this paper, we generated two mutants of Botrytis cinerea with AbA-resistance, BcAUR1a and BcAUR1b, through UV irradiation. BcAUR1a lost an intron and BcAUR1b had three amino acid mutations (L197P, F288S and T323A) in the AUR1 gene. AbA strongly inhibits the activity of IPC synthase in wild-type B. cinerea, which leads to distinct changes in cell morphology, including the delay in conidial germination, excessive branching near the tip of the germ tube and mycelium, and the inhibition of the mycelium growth. Further, AbA prevents the infection of wild-type B. cinerea in tomato fruits via reducing oxalic acid secretion and the activity of cellulase and pectinase. On the contrary, AbA has no effect on the growth and pathogenicity of the two mutants. Although both mutants show a similar AbA resistance, the molecular mechanisms might be different between the two mutants.
Asunto(s)
Antifúngicos/farmacología , Botrytis/enzimología , Botrytis/crecimiento & desarrollo , Depsipéptidos/farmacología , Hexosiltransferasas/genética , Hexosiltransferasas/metabolismo , Solanum lycopersicum/microbiología , Secuencia de Aminoácidos , Botrytis/efectos de los fármacos , Botrytis/patogenicidad , Depsipéptidos/antagonistas & inhibidores , Farmacorresistencia Fúngica Múltiple/genética , Proteínas Fúngicas/genética , Hexosiltransferasas/antagonistas & inhibidores , Mutación , Micelio/efectos de los fármacos , Micelio/crecimiento & desarrollo , Micelio/ultraestructura , Alineación de Secuencia , Esporas Fúngicas/efectos de los fármacos , Esporas Fúngicas/fisiología , Esporas Fúngicas/ultraestructuraRESUMEN
OBJECTIVE: To investigate the effect of nitric oxide (NO) on Toll-like receptors 2 and 4 (TLR 2/4) gene expression in the liver in acute hemorrhagic necrotizing pancreatitis (AHNP) in rats. METHODS: Seventy SD male rats were randomly divided into sham-operated group (n=10), AHNP group (n=30) and L-arginine (L-Arg) treatment group (n=30). Blood samples and liver tissues were obtained at 6 hours in sham-operated group, and 3, 6, 12 hours respectively in AHNP group and L-Arg-treated group. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), amylase in serum, NO, tumor necrosis factor-alpha (TNF-alpha) in liver tissue were determined. TLR2/4 mRNA expressions in the liver tissue were measured by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: TLR2/4 mRNA could be detected in the liver with low values in sham-operated group [(1.150+/-0.725)x10(-6), (11.450+/-1.724)x10(-4)], but they were markedly upregulated at 3 hours in AHNP group [(1.970+/-0.362)x10(-3), (175.000+/-0.111)x10(-3)], and peaked at 12 hours [(29.400+/-3.155)x10(-4), (267.300+/-8838)x10(-2), P<0.01). At the same time serum levels of amylase, ALT and AST increased, hepatic injuries were aggravated, the levels of TNF-alpha in the liver were increased and levels of NO in the liver were lowered (P<0.05 or P<0.01). Treatment with L-rg could effectively inhibit TLR2/4 mRNA expression [3 h: (3.510+/-1.528)x10(-4), (13.500+/-2.231)x10(-2); 6 h: (21.000+/-5.346)x10(-4), (18.700+/-2.685)x10(-2); 12 h: (26.200+/-2.076)x10(-4), 1.959+/-0.270, P<0.05 or P<0.01] and alleviate hepatic injuries. The levels of serum amylase, ALT and AST lowered, and the levels of TNF-alpha in the liver were lowered and the levels of NO were markedly increased (P<0.05 or P<0.01). CONCLUSION: These findings suggest that the expression of TLR2/4 mRNA is increased in the liver in AHNP, and the hepatic injuries are aggravated. NO could markedly inhibit TLR 2/4 mRNA gene expression in the liver in AHNP.
Asunto(s)
Óxido Nítrico/farmacología , Pancreatitis Aguda Necrotizante/metabolismo , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/metabolismo , Animales , Modelos Animales de Enfermedad , Hígado/metabolismo , Masculino , Pancreatitis Aguda Necrotizante/tratamiento farmacológico , ARN Mensajero/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
AIM: To investigate the role of nitric oxide (NO) in Toll-like receptor 2 (TLR2)/4mRNA expression in livers of acute hemorrhagic necrotizing pancreatitis (AHNP) rats. METHODS: One hundred and ten SD male rats were randomly divided into sham-operated group (n=10), AHNP group (n=30), chloroquine (CQ)-treated group (n=30) and L-Arg-treated group (n=40). TLR2/4mRNA expression in the liver of AHNP rats was measured by RT-PCR. RESULTS: Expression of TLR2/4mRNA could be detected in the liver of AHNP rats in sham-operated group (0.155E-5+/-0.230E-6 and 0.115E-2+/-0.545E-4), but was markedly increased at 3 h in AHNP group (0.197E-2+/-0.114E-3 and 0.175+/-0.349E-2) peaking at 12 h (0.294E-2+/-0.998E-4 and 2.673+/-2.795E-2, P<0.01). Hepatic injuries were aggravated, TNF-alpha concentration in the liver was increased and NO concentration was decreased (P<0.05 or P<0.01). When TLR2/4mRNA expression was inhibited by CQ (3 h: 1.037E-4+/-3.299E-6 and 0.026+/-3.462E-3; 6 h: 1.884E-4+/-4.679E-6 and 0.108+/-6.115E-3; 12 h: 2.443E-4+/-7.714E-6 and 0.348+/-6.807E-3; P<0.01), hepatic injuries were relieved, NO concentration in the liver was increased and TNF-alpha concentration was decreased (P<0.05 or P<0.01). When rats with AHNP were treated with L-Arg, TLR2/4mRNA expression in the liver could be effectively inhibited (50 mg-T: 0.232E-2+/-0.532E-4 and 0.230+/-6.883E-3; 100 mg-T: 0.210E-2+/-1.691E-4 and 0.187+/-0.849E-2; 200 mg-T: 0.163E-2+/-0.404E-4 and 0.107+/-0.195E-2; 400 mg-T: 0.100E-2+/-0.317E-4 and 0.084+/-0.552E-2; P<0.01) and hepatic injuries were relieved. At the same time, NO concentration in the liver was markedly increased and TNF-alpha concentration was decreased (P<0.05 or P<0.01). CONCLUSION: The expression of TLR2/4mRNA is increased and hepatic injuries are aggravated in the liver of AHNP rats. TLR2/4mRNA gene expression in the liver of AHNP rats can be markedly inhibited by NO, leading to the relief of hepatic injuries.
Asunto(s)
Hemorragia/metabolismo , Hígado/metabolismo , Óxido Nítrico/metabolismo , Pancreatitis Aguda Necrotizante/metabolismo , ARN Mensajero/metabolismo , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/metabolismo , Alanina Transaminasa/sangre , Amilasas/sangre , Animales , Aspartato Aminotransferasas/sangre , Humanos , Masculino , ARN Mensajero/genética , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptor Toll-Like 2/genética , Receptor Toll-Like 4/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: Toll-like receptor (TLR) 2/4 might play important roles in mediating proinflammatory cytokine synthesis and release. And nitric oxide (NO) has been used to treat acute respiratory distress syndrome (ARDS). This study aimed to investigate the changes in TLR2/4 gene expression in the lungs of rats with acute lung injury (ALI) complicated by acute hemorrhage necrotizing pancreatitis (AHNP) and the effect of NO on the TLR2/4 gene expression. METHODS: One hundred and ten SD male rats were randomly divided into sham-operated group (n=10), AHNP group (n=30), chloroquine-treated group (n=30), and L-Arg-treated group (n=40). The lungs were dissected for lung histological scoring, and bronchoalveolar lavages were harvested for lung injury indexing. TLR2/4 mRNA expression in the lungs was measured by RT-PCR. RESULTS: TLR2/4mRNA was detected in the lungs with low values in the sham-operated group (0.016+/-0.210E-2, 0.112+/-0.750E-2), but it was markedly increased at 3 hours in the AHNP group (0.787+/-0.751E-2, 1.512+/-1.794E-2), peaking at 12 hours (1.113+/-6.141E-2, 2.957+/-2.620E-2; P<0.05 or P<0.01). When lung injuries were aggravated, TNF-alpha concentrations in the lungs were increased, but NO concentrations were decreased (P<0.05 or P<0.01). When TLR2/4mRNA was inhibited by CQ (3h: 0.313+/-5.491E-2, 0.005+/-1.419E-3; 6h: 0.488+/-7.442E-2, 0.010+/-1.518E-3; 12h: 0.883+/-8.911E-2, 0.024+/-2.760E-3; P<0.05 or P<0.01), lung injuries were relieved. NO concentrations in the lungs were increased but TNF-alpha concentrations were decreased (P<0.05 or P<0.01). When the rats with AHNP were treated with L-Arg, TLR2/4mRNA expression in the lungs could be effectively inhibited (50 mg-T: 0.656+/-3.977E-2, 1.501+/-6.111E-2; 100 mg-T: 0.260+/-0.891E-2, 0.732+/-5.135E-2; 200 mg-T: 0.126+/-0.914E-2, 0.414+/-1.678E-2; 400 mg-T: 0.091+/-0.399E-2, 0.287+/-0.176E-2;P<0.05 or P<0.01) and lung injuries were relieved. At the same time, NO concentrations in the lungs were markedly increased, but TNF-alpha concentrations were decreased (P<0.05 or P<0.01). CONCLUSIONS: The expression of TLR2/4mRNA is increased in the lungs in rats with AHNP and lung injuries are aggravated.TLR2/4mRNA gene expression of the lungs of rats with AHNP could be markedly inhibited by NO, leading to the relief of lung injuries.