RESUMEN
In the tumor progression, transforming growth factor ß1 (TGFß1) plays a critical role in tumorigenesis as well as metastasis. It is known that high plasma level of TGFß1 in patients with advanced non-small cell lung cancer (NSCLC) is correlated with poor prognostics. In addition, the generation of cancer stem-like cells is associated with metastasis, drug resistance, and tumor recurrence, which also lead to poor outcomes in NSCLC patients. However, it remains unclear how TGFß1 promotes NSCLC cells to acquire stem-like properties and accelerate tumor metastasis. In our study, we found that short term TGFß1 treatment resulted in a significant epithelial-mesenchymal transition (EMT) morphological change in TGFß1-sensitive NSCLC cells but not in insensitive cells. Western blotting confirmed increased Vimentin and reduced E-Cadherin protein expression after TGFß1 treatment in A549, NCI-H1993, and NCI-H358 cells. TGFß1 incubation dramatically decreased in vitro cell proliferation and increased cell invasion in TGFß1-sensitive NSCLC cells but not in NCI-H1975, NCI-H1650, and HCC827 cells. Moreover, TGFß1 was able to enhance the mRNA expression of Oct4, Nanog and Sox2 and drastically increased anchorage-independent colony formation in TGFß1-sensitive NSCLC cells, suggesting the acquisition of cancer stem-like properties. Interestingly, we found that vascular endothelial growth factor receptor 3 (VEGFR3) mRNA expression was significantly elevated in TGFß1-sensitive NSCLC cells compared to insensitive cells. And TGFß1 was capable of inducing VEGF-C gene expression. Pharmacological blocking TGFß type I receptor kinase (ALK5) significantly inhibited TGFß1-induced VEGF-C expression. Silencing of ALK5 by siRNA also dramatically reduced TGFß1-induced VEGF-C expression in TGFß1-sensitive NSCLC cells. Therefore, TGFß1 contributes for NSCLC metastasis through promoting EMT, generation of high invasive cancer cells with stem-like properties, and increasing VEGF-C expression. Blocking TGFß pathway is a potential therapeutic target in human non-small cell lung cancer.
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Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Transición Epitelial-Mesenquimal/fisiología , Neoplasias Pulmonares/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Factor C de Crecimiento Endotelial Vascular/metabolismo , Receptor 3 de Factores de Crecimiento Endotelial Vascular/metabolismo , Cadherinas/metabolismo , Línea Celular Tumoral , Proliferación Celular/fisiología , Regulación Neoplásica de la Expresión Génica , Humanos , Invasividad Neoplásica/fisiopatología , Células Madre Neoplásicas/metabolismo , ARN Mensajero/metabolismo , Receptor Tipo I de Factor de Crecimiento Transformador beta/antagonistas & inhibidores , Receptor Tipo I de Factor de Crecimiento Transformador beta/metabolismo , Factor de Crecimiento Transformador beta1/administración & dosificación , Vimentina/metabolismoRESUMEN
Abnormal microRNA-370 (miR-370) expression has been frequently reported in several types of cancers, including lung cancer. However, the role and molecular mechanisms of miR-370 in regulating the growth and metastasis of lung cancer have not been clarified. Here, we show higher levels of epidermal growth factor receptor (EGFR), but lower levels of miR-370 expression in most human lung cancer cells and non-tumor cells. Induction of miR-370 over-expression significantly reduced the levels of EGFR expression and the EGFR 3'untranslated region (UTR)-regulated luciferase activity in XWLC-05 and H157 cells, suggesting that miR-370 may bind to the 3'UTR of EGFR mRNA. Compared with the control cells, induction of miR370 overexpression significantly inhibited the proliferation, clone formation capacity, migration and invasion of XWLC-05 and H157 cells while miR-370 inhibitor over-expression enhanced their tumor behaviors in vitro. Furthermore, miR-370 over-expression down-regulated the EGFR and hypoxia-inducible factor (HIF)-1α expression, and attenuated the extracellular single-regulated kinase (ERK)1/2 and AKT phosphorylation in XWLC-05 and H157 cells. In contrast, miR370 inhibitor over-expression increased the EGFR and HIF-1α expression as well as the ERK1/2 and AKT phosphorylation in XWLC-05 and H157 cells. Moreover, miR-370 over-expression significantly reduced the levels of EGFR and CD31 expression and inhibited the growth and lung metastasis of xenograft NSCLC tumors in mice. Our study indicates that miR-370 may bind to the 3'UTR of EGFR to inhibit EGFR expression and the growth, angiogenesis and metastasis of non-small cell lung cancer by down-regulating the ERK1/2 and AKT signaling.
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Bone is one of the most preferred sites of metastasis in lung cancer. Currently, bisphosphonates and denosumab are major agents for controlling tumor-associated skeletal-related events (SREs). However, both bisphosphonates and denosumab significantly increase the risk for jaw osteonecrosis. Statins, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors and the most frequently prescribed cholesterol-lowering agents, have been reported to inhibit tumor progression and induce autophagy in cancer cells. However, the effects of statin and role of autophagy by statin on bone metastasis are unknown. In this study, we report that fluvastatin effectively prevented lung adenocarcinoma bone metastasis in a nude mouse model. We further reveal that fluvastatin-induced anti-bone metastatic property was largely dependent on its ability to induce autophagy in lung adenocarcinoma cells. Atg5 or Atg7 deletion, or 3-methyadenine (3-MA) or Bafilomycin A1 (Baf A1) treatment prevented the fluvastatin-induced suppression of bone metastasis. Furthermore, we reveal that fluvastatin stimulation increased the nuclear p53 expression, and fluvastatin-induced autophagy and anti-bone metastatic activity were mostly dependent on p53.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Antineoplásicos/uso terapéutico , Neoplasias Óseas/prevención & control , Ácidos Grasos Monoinsaturados/uso terapéutico , Indoles/uso terapéutico , Neoplasias Pulmonares/tratamiento farmacológico , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Adenocarcinoma del Pulmón , Animales , Antineoplásicos/farmacología , Autofagia/efectos de los fármacos , Neoplasias Óseas/metabolismo , Neoplasias Óseas/secundario , Línea Celular Tumoral , Ácidos Grasos Monoinsaturados/farmacología , Femenino , Fluvastatina , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Indoles/farmacología , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Ratones Endogámicos BALB C , Ratones Desnudos , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
OBJECTIVE: The aim of this study was to investigate the correlation between the efficacy of epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) and circulating tumor cell (CTC) levels in patients with advanced non-small cell lung cancer (NSCLC). The efficacy of EGFR-TKIs in reducing CTC counts in patients with advanced NSCLC was studied. PATIENTS AND METHODS: A total of 66 patients with advanced NSCLC were enrolled and divided into two groups (those with high CTC counts and those with low CTC counts) based on the patients' median CTC counts. All the patients were treated with an EGFR-TKI, and the treatment efficacy and prognoses were compared. RESULTS: The treatment efficacies were 53.3% (16/30) and 27.8% (10/36) for the low CTC group and high CTC group, respectively, and this difference was statistically significant (P<0.05). The median overall survival was 22.8 months (95% confidence interval [CI]: 18.9-26.8 months) for the low CTC group and 18.3 months (95% CI: 2.9-8.2 months) for the high CTC group. The median progression-free survival was 11.5 months (95% CI: 8.1-15 months) and 5.6 months (95% CI: 2.9-8.2 months) for the low and high CTC groups, respectively, and the difference was statistically significant (P<0.05). CONCLUSION: The CTC count can be used as an index for predicting the EGFR-TKI effect on patients with advanced NSCLC. Efficacy and prognosis of EGFR-TKI treatment and CTC count were considered important, and the CTC count could be used to predict the efficacy of EGFR-TKI treatment and prognosis of advanced NSCLC. The change in CTC expression levels can be used as an index for evaluating the prognosis of patients with advanced NSCLC.
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Heat shock protein 90 (HSP90) is a molecular chaperone required for the stability and function of many proteins. The chaperoning of oncoproteins by HSP90 enhances the survival, growth, and invasive potential of cancer cells. HSP90 inhibitors are promising new anticancer agents, in which the benzoquinone ansamycin 17-allylamino-17-demethoxygeldanamycin (17-AAG) is currently in clinical evaluation. However, the implications of acquired resistance to this class of drug remain largely unexplored. In the present study, we have generated isogenic human colon cancer cell lines that are resistant to 17-AAG by continued culturing in the compound. Cross-resistance was found with another HSP90 inhibitor 17-dimethylaminoethylamino-17-demethoxygeldanamycin. The resistant cells showed obvious morphology changes with a metastatic phenotype and significant increases in migration and adhesion to collagens. Western blotting analysis of epithelial-mesenchymal transition molecular markers found that expression of E-cadherin downregulated, whereas expression of N-cadherin and ß-catenin upregulated in the resistant cells. Mucin 1 (MUC1) has been reported to mediate metastasis as well as chemical resistance in many cancers. Here, we found that MUC1 expression was significantly elevated in the acquired drug resistance cells. 17-AAG treatment could decrease MUC1 more in parental cells than in acquired 17-AAG-resistant cells. Further study found that knockdown of MUC1 expression by small interfering RNA could obviously re-sensitize the resistant cells to 17-AAG treatment, and decrease the cell migration and adhesion. These were coupled with a downregulation in N-cadherin and ß-catenin. The results indicate that HSP90 inhibitor therapies in colon carcinomas could generate resistance and increase metastatic potential that might mediated by upregulation of MUC1 expression. Findings from this study further our understanding of the potential clinical effects of HSP90-directed therapies in colon carcinomas.
Asunto(s)
Antineoplásicos/farmacología , Benzoquinonas/farmacología , Neoplasias del Colon/patología , Resistencia a Antineoplásicos , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Lactamas Macrocíclicas/farmacología , Mucina-1/metabolismo , Adhesión Celular , Línea Celular Tumoral/efectos de los fármacos , Movimiento Celular , Neoplasias del Colon/metabolismo , Técnicas de Inactivación de Genes , Humanos , Mucina-1/genética , Metástasis de la Neoplasia , Interferencia de ARNRESUMEN
Indoleamine 2, 3-dioxygenase (IDO) is a rate-limiting enzyme for tryptophan metabolism inducing immune tolerance of tumors. The purpose of this study is to investigate IDO expression and its prognostic significance in bladder urothelial carcinoma (BUC). In this study, immunohistochemical staining for IDO expression in BUC tissues (n = 84) and normal bladder tissues (n = 22) was performed. The mRNA expression levels of IDO in BUC and normal bladder were analyzed by quantitative RT-PCR. Survival analysis was performed for the correlation of IDO expression and clinicopathological factors with disease-free survival. Positive expression of IDO was found in 48 of 84 cases in BUC tissues and was significantly correlated with histological classification, histological grade and TNM stage. While IDO expression in normal bladder tissues was expressed in only 4 of 22 (18.2%) cases. Moreover, IDO mRNA levels of BUC were significantly higher than that of normal bladder. We also found that IDO, histological grade and TNM stage were closely associated with DFS. These results indicated that IDO was related to the progression of BUC and might be one of the crucial prognostic factors for BUC.
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Biomarcadores de Tumor/análisis , Carcinoma/enzimología , Indolamina-Pirrol 2,3,-Dioxigenasa/análisis , Neoplasias de la Vejiga Urinaria/enzimología , Urotelio/enzimología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/genética , Carcinoma/genética , Carcinoma/mortalidad , Carcinoma/patología , Carcinoma/cirugía , Progresión de la Enfermedad , Supervivencia sin Enfermedad , Femenino , Humanos , Inmunohistoquímica , Indolamina-Pirrol 2,3,-Dioxigenasa/genética , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Estadificación de Neoplasias , ARN Mensajero/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/mortalidad , Neoplasias de la Vejiga Urinaria/patología , Neoplasias de la Vejiga Urinaria/cirugía , Urotelio/patología , Urotelio/cirugía , Adulto JovenRESUMEN
The transcription factor SOX4 has functional importance in foetal lung maturation and tumorigenesis in a number of cancers. However, its biological functions in the progression of lung tumorigenesis remain unclear. In this study, we found that the expression levels of SOX4 mRNA and protein were significantly higher in Xuanwei female lung cancer tissues than in benign lung lesions. The patients with high expression of the SOX4 protein had a higher pathological grade, lymph node (LN) metastasis, poor tumor differentiation and worse prognosis than those patients with low expression of SOX4. Knockdown of the SOX4 gene in the Xuanwei female lung cancer cell line XWLC-05 resulted in apoptotic morphological changes, decreased cell proliferation, invasion and migration. Furthermore, knockdown of the SOX4 gene resulted in obvious sub-G1 peaks and induction of apoptosis through upregulation of caspase-3 expression, while in cells treated with a caspase-3 inhibitor, apoptosis induced by silencing SOX4 expression was inhibited. In vivo analysis in nude mice further confirmed that knockdown of SOX4 suppressed tumor growth. In conclusion, SOX4 appears to be an important tumor suppressor gene in the regulation of Xuanwei female lung cancer cell proliferation, apoptosis and metastases, and it may be a potential target for effective lung cancer therapy.
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Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Factores de Transcripción SOXC/metabolismo , Animales , Apoptosis/genética , Apoptosis/fisiología , Western Blotting , Proliferación Celular/fisiología , Femenino , Humanos , Inmunohistoquímica , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia , Metástasis Linfática/genética , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Microscopía Electrónica de Transmisión , Persona de Mediana Edad , Reacción en Cadena en Tiempo Real de la Polimerasa , Factores de Transcripción SOXC/genéticaRESUMEN
AIMS: To investigate changes in cellular immune function of patients with lung cancer before and after cytokine- induced killer (CIK) cell therapy and to identify variation effects on overall survival (OS) and progression-free survival (PFS). MATERIALS AND METHODS: A total of 943 lung cancer patients with immune dysfunction were recruited from January 2002 to January 2010, 532 being allocated to conventional therapy and 411 to CIK therapy after a standard treatment according to the NCCN Clinical Practice Guidelines. All the patients were investigated for cellular immune function before and after therapy every three months. and clinical prognostic outcomes were analyzed. RESULTS: After six courses of treatment, immune function was much improved in patients receiving CIK cells therapy as compared to controls. The percentages of recurrence and/or metastases for patients undergoing CIK cell therapy was 56.2% and 49.1% respectively but 78.6% and 70.3% among controls (p<0.001). The median OS times for CIK cell therapy and control groups were 48 and 36 months respectively. The OS rates at 12, 36, 60, 84 months in CIK treated patients were 97.8%, 66.9%, 27.7%, and 4.1% while they were 92.3%, 44.5%, 9.2%, and 1.5% in controls. OS and PFS were significantly different by log rank test between the two groups and across the three immune improvement classes. CONCLUSIONS: The immune function of lung cancer patients was improved by CIK cell therapy, associated with an increase in the OS rate and extension of the time to recurrence and/or metastasis.
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Adenocarcinoma/terapia , Carcinoma de Pulmón de Células no Pequeñas/terapia , Carcinoma de Células Escamosas/terapia , Tratamiento Basado en Trasplante de Células y Tejidos , Células Asesinas Inducidas por Citocinas/inmunología , Neoplasias Pulmonares/terapia , Recurrencia Local de Neoplasia/terapia , Adenocarcinoma/inmunología , Adenocarcinoma/mortalidad , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Carcinoma de Células Escamosas/inmunología , Carcinoma de Células Escamosas/mortalidad , Estudios de Casos y Controles , Femenino , Estudios de Seguimiento , Humanos , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/mortalidad , Metástasis Linfática , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Recurrencia Local de Neoplasia/inmunología , Recurrencia Local de Neoplasia/mortalidad , Estadificación de Neoplasias , Pronóstico , Tasa de SupervivenciaRESUMEN
In this study, a central venous catheter (CVC)-associated infection model was established in rats to investigate and evaluate the effect of biofilms on the virulence of the pathogens. Twenty-four adult SD rats were randomly divided into biofilm positive (BF+) and biofilm negative (BF-) groups to be challenged with strains of S.epidermidis. Serum levels of inflammatory cytokines were measured and the infection rate and counts of bacteria cells were studied. Compared to rats of BF- group, the serum level of TNF and IL-6 significantly increased in rats of BF+ group (P < 0.01) and the level of IL-10 and IFN-γ significantly decreased (P < 0.01), striking the balance of pro-inflammatory/anti-inflammatory cytokines. The infection rate and bacterial counts in tissues and blood of rats of BF + group were significantly higher than those of rats of BF- group (P < 0.05).Inflammatory cell infiltration in vital organs (heart, lung, liver and kidneys) was more significant in rats of BF+ group than that of rats of BF- group. CVC-associated infection model can be successfully reproduced in rats by injecting 5 × 10(6) CFU of S.epidermidis. Biofilm formation can significantly enhance the virulence of the bacteria, leading to uncontrolled infection. The serum level of inflammatory cytokines, infection rate and the extent of inflammatory cell infiltration are important markers for evaluating the virulence of biofilm.
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Biopelículas/crecimiento & desarrollo , Infecciones Relacionadas con Catéteres/etiología , Catéteres Venosos Centrales/efectos adversos , Modelos Animales de Enfermedad , Infecciones Estafilocócicas/etiología , Staphylococcus epidermidis/fisiología , Animales , Infecciones Relacionadas con Catéteres/sangre , Catéteres Venosos Centrales/microbiología , Citocinas/sangre , Masculino , Especificidad de Órganos , Ratas , Infecciones Estafilocócicas/sangre , Staphylococcus epidermidis/patogenicidad , VirulenciaRESUMEN
Gold nanorods (AuNRs) have been used in plasmonic photothermal therapy (PPTT), which is thought to be more efficient and selective than conventional photothermal therapy. The efficiency and safety of PPTT can be improved by functionally modifying the gold nanorods with proteins or biomolecules. In this study, AuNRs were modified with anti-epidermal growth factor receptor (EGFR) monoclonal antibody (mAb), and the apoptotic potential of EGFRmAb-AuNR was assessed in Hep-2 cells in vitro and in vivo. The EGFRmAb modification had no obvious influence on the original optical property of the AuNRs, but it significantly increased the entry of AuNRs into Hep-2 cells. EGFRmAb-AuNRs, with appropriate laser irradiation, resulted in higher Hep-2 cells apoptosis than AuNRs did alone, in vitro, and was accompanied by alteration of reactive oxygen species (ROS) production, Ca(2+) release, change in mitochondrial membrane potential (ΔΨm), cytochrome c (Cyt-c) release, active caspase-3 expression, and level of B-cell lymphoma 2 (Bcl-2) and B-cell lymphoma 2 protein-associated X protein (Bax). EGFRmAb-AuNR-mediated apoptosis in Hep-2 cells was also observed in vivo and had an inhibitive effect on growth of Hep-2 tumor xenografts. Our data suggest that the EGFRmAb modification improves AuNR-mediated apoptosis and may have the potential to be used clinically.
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Anticuerpos Monoclonales/inmunología , Apoptosis/efectos de los fármacos , Receptores ErbB/inmunología , Oro/uso terapéutico , Nanopartículas del Metal/uso terapéutico , Nanocompuestos/uso terapéutico , Fototerapia/métodos , Animales , Anticuerpos Monoclonales/química , Células Hep G2 , Humanos , Nanopartículas del Metal/química , Nanopartículas del Metal/ultraestructura , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Nanocompuestos/química , Nanocompuestos/ultraestructura , Nanotubos/química , Nanotubos/ultraestructura , Tamaño de la Partícula , Fármacos Fotosensibilizantes , Resultado del TratamientoRESUMEN
OBJECTIVE: To study the effect of targeted Sox4 gene-knock-down on the growth of xenografts of Xuanwei female lung cancer cell line XWLC-05 cells in nude mice. METHODS: Recombinant plasmid pGFP-V-RS-Sox4 shRNA was constructed and transfected into XWLC-05 cells. Real-time quantitative PCR and Western blot were applied to confirm the effect of Sox4 gene-knock-down. XWLC-05 cells stably transfected with the plasmids were inoculated into nude mice to establish the xenograft model. The nude mouse status, tumor formation and tumor growth were observed, and the tumor inhibition rate was calculated. CT scan was performed to assess the metastasis of xenografts. Immunohistochemical staining was applied to detect Sox4 and ki-67 protein expression. RESULTS: Recombinant plasmid pGFP-V-RS-A-Sox4 shRNA which can effectively knocking-down Sox4 gene was successfully constructed and the stable transfected cells were selected by puromycin-screening. The success rate of tumor cell inoculation was 100% in the mice of all groups except those inoculated with saline. The body weight of all mice inoculated with parental XWLC-05 cells (blank control), pGFP-V-RS-scram shRNA trsfected XWLC-05 cells (negative control), and pGFP-V-RS-Sox4 shRNA transfected XWLC-05 cells was increased to a varying degree, but there was no significant difference among the groups (P > 0.05 ). The growth of xenografts was significantly inhibited after silencing the Sox4 gene expression when compared with that of the blank and negative controls (P < 0.05) . The volume of removed tumors of the Sox4 gene-inhibited mice was (2.30 ± 0.34) cm(3) , significantly smaller than that of the negative control (3.99 ± 0.45) cm(3) and the blank control (4.03 ± 0.42) cm(3) (P < 0.05) . The weight of removed tumors of Sox4 gene-inhibited mice was (0.86 ± 0.14) g, significantly lower than that of the negative control (1.84 ± 0.27) g and blank control (1.86 ± 0.22) g, (P < 0.05). Immunohistochemical staining showed that Sox4 and ki-67 proteins mainly expressed in cell nuclei. The staining was significantly decreased in xenografts of Sox4-inhibited mice when compared with the negative and blank controls (P < 0.05). No distant metastasis was found in any mouse by CT imaging and pathological examination during the observation period. CONCLUSIONS: The xenograft model of Xuanwei female lung cancer cell line XWLC-05 cells in nude mice is successfully established. Knocking-down of Sox4 gene can suppress the xenograft tumor growth.
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Técnicas de Silenciamiento del Gen , Neoplasias Pulmonares/genética , Factores de Transcripción SOXC/metabolismo , Animales , Femenino , Xenoinjertos , Humanos , Neoplasias Pulmonares/patología , Ratones , Ratones DesnudosRESUMEN
Non-small cell lung cancer (NSCLC) is the most common cause of cancer-related death in both men and women worldwide. Recently, Disulfiram has been reported to be able to inhibit glioblastoma, prostate, or breast cancer cell proliferation. In this study, the synergistic effect of Disulfiram and copper on NSCLC cell growth was investigated. Inhibition of cancer cell proliferation was detected by 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) assay and cell cycle analysis. Liquid colony formation and tumor spheroid formation assays were used to evaluate their effect on cancer cell clonogenicity. Real-time PCR was performed to test the mRNA level of cancer stem cell related genes. We found that Disulfiram or copper alone did not potently inhibit NSCLC cell proliferation in vitro. However, the presence of copper significantly enhanced inhibitory effect of Disulfiram on NSCLC cell growth, indicating a synergistic effect between Disulfiram and copper. Cell cycle analysis showed that Disulfiram/copper complex caused NSCLC cell cycle arrest in G2/M phase. Furthermore, Disulfiram/copper significantly increased the sensitivity of cisplatin in NSCLC cells tested by MTT assay. Liquid colony formation assay revealed that copper dramatically increased the inhibitory effect of Disulfiram on NSCLC cell colony forming ability. Disulfiram combined with copper significantly attenuated NSCLC cell spheroid formation and recuded the mRNA expression of lung cancer stem cell related genes. Our data suggest that Disulfiram/copper complex alone or combined with other chemotherapy is a potential therapeutic strategy for NSCLC patients.
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Antineoplásicos/farmacología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Cobre/farmacología , Disulfiram/farmacología , Inhibidores Enzimáticos/farmacología , Neoplasias Pulmonares/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/patología , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cisplatino/farmacología , Sinergismo Farmacológico , Humanos , Pulmón/efectos de los fármacos , Pulmón/patología , Neoplasias Pulmonares/patologíaRESUMEN
AIMS: To study the CIK cell treatment effects on regulation of cellular immune function disorders in patients with lung cancer, and to analyze the time characteristics. METHODS: Cellular immune function was assessed by FCM, and patients with functional disorders were randomly divided into two groups, one given CIK cell therapy within 18 months (5 courses) and the other the controls, which were followed up for 1 year with cellular immune functions tested once a month. RESULTS: There were 5 types of cellular immunity, 4 of which are disorders; after CIK treatment, the improvement rate of the 4 groups were 79.1%, 70.8%, 76.0% and 70.0%, intergroup differences not being statistically significant (P=0.675), all significantly higher than in the control group (P=0.000). The median maintenance times for the 4 groups were 10.4 months (9.76-11.04), 8.4 months (7.86-8.94), 9.8 months (9.20-10.4) and 7.9 months (6.25-9.55), respectively. CONCLUSIONS: CIK cells were able to improve the immune functions of patients with lung cancer, the rate of improvement and maintenance time being related to the immune function before the treatment and CIK-cell-therapy courses.
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Tratamiento Basado en Trasplante de Células y Tejidos , Células Asesinas Inducidas por Citocinas/inmunología , Neoplasias Pulmonares/terapia , Pulmón/inmunología , Adulto , Anciano , Estudios de Casos y Controles , Femenino , Estudios de Seguimiento , Humanos , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/mortalidad , Masculino , Persona de Mediana Edad , Pronóstico , Tasa de Supervivencia , Adulto JovenRESUMEN
OBJECTIVES: Lymph node involvement is a key feature and an independent prognostic factor of oesophageal squamous cell carcinoma. However, an accurate and robust assay to predict the lymphatic spread of oesophageal squamous cell carcinoma is unavailable. The purpose of this study was to determine whether serum vascular endothelial growth factor-C (VEGF-C) and spleen tyrosine kinase levels are potential markers of lymph node metastasis in patients with oesophageal squamous cell carcinoma. METHODS: In the study, 108 patients with oesophageal squamous cell carcinoma and 24 healthy subjects participated. Serum spleen tyrosine kinase and VEGF-C concentrations were examined by enzyme-linked immunosorbent assays. RESULTS: Patients with oesophageal squamous cell carcinoma had significantly elevated VEGF-C and decreased spleen tyrosine kinase in serum when compared with normal controls (P < 0.05). Pearson's correlation analysis revealed that serum spleen tyrosine kinase was negatively correlated with the serum VEGF-C level in oesophageal squamous cell carcinoma patients (r = -0.453, P = 0.000). In addition, surgical resections of the oesophageal tumour profoundly brought both serum spleen tyrosine kinase and VEGF-C closer to the normal range at 2 weeks after operation (P < 0.05). Statistical analysis showed that enhanced serum VEGF-C and reduced spleen tyrosine kinase significantly correlated with the stage of regional lymph node metastasis, tumour size and the status of primary tumour extension, but not with other clinicopathological features of the patients (P < 0.05). CONCLUSIONS: Our data suggest that both serum spleen tyrosine kinase and VEGF-C levels are valuable in the preoperative evaluation of lymph node metastasis in patients with oesophageal squamous cell carcinoma.
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Carcinoma de Células Escamosas/sangre , Carcinoma de Células Escamosas/patología , Neoplasias Esofágicas/sangre , Neoplasias Esofágicas/patología , Péptidos y Proteínas de Señalización Intracelular/sangre , Proteínas Tirosina Quinasas/sangre , Factor C de Crecimiento Endotelial Vascular/sangre , Anciano , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Curva ROC , Quinasa SykRESUMEN
BACKGROUND: The incidence of Xuanwei lung cancer ranks first in China, and its pathogenesis requires in-depth investigation. This study aims to establish an orthotopic Xuanwei lung cancer severe combined immunodeficiency (SCID) mouse model and to provide a basic experimental platform for further study. METHODS: The Xuanwei lung cancer cell line XWLC-05 was inoculated into the lung tissue of SCID mice in high and low doses. The tumor formation rates, tumor characteristics, spontaneous metastases, and survival times of the mice were observed, taking a subcutaneously transplanted tumor as control. RESULTS: The tumor formation rates of the orthotopic transplantation of lung cancer cells in high and low doses were 81% and 83%, respectively, among which mice in the high-dose group appeared cachectic on day 13. Extensive invasion and adhesion were observed in the contralateral lung and thoracic cavity, but no distant metastasis was exhibited. Mice with low-dose cells in the orthotopic transplantation group appeared cachectic and distant metastasis occurred on day 25. The tumor formation rates in the subcutaneous inoculation group by the high and low doses of cells were 100% and 94.5%, respectively, and no distant metastasis was observed. The rate of metastasis within the orthotopic transplantation group and between the orthotopic and subcutaneous inoculation groups showed a significant difference (P<0.05). A significant difference was indicated by the survival rate within and between the groups (P<0.001). CONCLUSIONS: We successfully established an orthotopic XWLC SCID mouse model, which lays the foundation for a more in-depth study.
Asunto(s)
Transformación Celular Neoplásica , Modelos Animales de Enfermedad , Neoplasias Pulmonares/patología , Animales , Carcinogénesis , Línea Celular Tumoral , Femenino , Masculino , Ratones , Ratones SCID , Metástasis de la Neoplasia , Análisis de SupervivenciaRESUMEN
Brachytherapy is regarded as the most effective method in the treatment of metastatic spinal tumors since little damage is caused to surrounding healthy tissue. However, this method may cause radiation myelopathy if an overdose occurs. In the present study, we established a Banna mini-pig (125)I spinal cord implantation model to provide a tool for the study of how to reduce these types of side effects. Cell cycle alteration, apoptosis and necrosis of spinal cord neurons in the presence of various doses and durations of (125)I brachytherapy were also investigated. The pigs were randomly divided into four groups, A, B, C and D. In group A, four (125)I seeds (total radioactivity, 4.0 mCi) were implanted into the dura mater of the spinal canal at the level of T13. In groups B and C, eight (125)I sources (total radioactivity, 8.0 mCi) were inserted at the same location. Groups A and C were raised for up to 8 months and group B for only 2 months. Neurons from the swine spinal cord at the T13 level were collected and cell cycle analysis was performed. Apoptosis and necrosis were tested by a terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay. The Banna mini-pig brachytherapy model was successfully established. Radiation myelopathy was closely associated with radiation dose and duration, more neurons were blocked in the G2 and S phases as dose and time increased, and an increase in apoptosis and necrosis was detected. Ratios of apoptosis and necrosis were reduced as lower doses and shorter durations of radiation were applied. Our results demonstrate that the Banna mini-pig is an ideal animal to study (125)I brachytherapy. Low-dose and short-term brachytherapy may effectively decrease apoptosis and necrosis in spinal cord cells in Banna mini-pigs.
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BACKGROUND: Selenium is an essential micronutrient for mammals but toxic in large amounts. Most studies indicate that selenium has inhibitory effect on cancer. The aim of this study is to investigate the effects of selenium and reduced glutathione (GSH) combined application on the proliferation and apoptosis of human lung adenocarcinoma cell line XWLC-05. METHODS: XWLC-05 cells were respectively treated in vitro by four factors (sodium selenite, GSH, sodium selenite+GSH and blank control (RMPI-1640 +10% calf serum) in different concentrations for 24 h. Cell growth inhibition rates were determined by MTT assay, cytomorphology was observed under inverted phase contrast microscope and changes of cell cycle were detected by Flow Cytometry (FCM). RESULTS: Both selenium and GSH individual on the XWLC-05 cells were found to possess obvious growth inhibition effect on the XWLC-05 cells. Selenium and GSH combined application on the XWLC-05 cells had cooperative inhibition effect (P <0.01). The inhibition rate was increased in a dose-dependent relationship as selenium with concentrations between (0.5-4.0) mug/mL (P <0.01) whether it was selenium single factors or selenium and GSH combined effect. FCM results showed that some XWLC-05 cells were induced apoptosis and G1 phase cells were markedly increased and S, G2/M phase cells decreased in both selenium individual groups and selenium and GSH combined groups. CONCLUSIONS: Selenium and GSH combined application on XWLC-05 cells can enhance directly the cell growth inhibitory effect compared with selenium and GSH individual. The mechanism seems to inhibit the synthesis of RNA and protein and prevent cells from entering S phase.
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BACKGROUND: It has been known that vascular endothelial growth factor (VEGF) and its receptor (VEGFR2) play important roles in tumor angiogenesis. The aim of this study is to investigate whether an oral DNA vaccine against VEGFR2 has the inhibition effect on tumor growth and angiogenesis, and explore its mechanism in vivo. METHODS: C57BL/6 mice were respectively given the DNA vaccine encoding VEGFR2 (vaccine group), pcDNA3.1 (plasmid group) and saline (saline group). All the mice were then inoculated with Lewis lung carcinoma 3LL cells. Weight, size and microvessel density (MVD) of transplanted tumors were observed. The levels of CD3+ and CD8+ T cells in peripheral blood of mice were detected by flow cytometry. RESULTS: Weight of transplanted tumors in vaccine group was significantly smaller than those in plasmid and saline groups (P < 0.05), and MVD was significantly lower in vaccine group than that in plasmid and saline groups (P < 0.05). After inoculated with 3LL cells, CD3+ and CD8+ T cell levels of vaccine group were markedly higher than those of plasmid and saline groups (P < 0.05). CONCLUSIONS: The oral DNA vaccine can significantly inhibit angiogenesis and growth of transplanted tumor in mice. It may act through killing endothelial cells of tumor.
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BACKGROUND: It has been known that Kangfuxin, a drug derived from Periplaneta Americana, can induce cell apoptosis of many cancer cell lines in vitro. The aim of this study is to investigate the inhibitory effect and mechanism of Periplaneta Americana extract (PAE) on 3LL lung cancer in mice. METHODS: The C57BL/6J mice transplanted with 3LL lung cancer were divided into normal saline (NS), PAE high dose (PAE-H) and PAE low dose (PAE-L) groups. The body weight changes and inhibitory rate of tumor growth in each group were observed. In addition, the cell cycle, apoptosis index (AI) and the expression of apoptosis associated genes were analysed by flow cytometry (FCM). RESULTS: The body weights were decreased in PAE-L and PAE-H treated group compared with NS group and the inhibitive rate of tumor growth was 41.24% and 81.08% respectively. FCM assay indicated that PAE could induce apoptosis of lung cancer cell, and the apoptosis rate was concentration-dependent. At the same time, the number of S and G2/M phase cells was decreased, most of the cells were arrested in G1/G1 phase. The result of TUNEL showed that there were apoptosis and necrosis associated with upregulated expression of Fas, FasR and p53 genes, and downregulated expression of Bcl-2. CONCLUSIONS: PAE may inhibit the growth of 3LL lung cancer in mice and induce apoptosis of 3LL lung cancer cells. It might be related to its effects on the regulation of apoptotic gene expression.