RESUMEN
The Ras proteins play an important role in cell growth, differentiation, proliferation and survival by regulating diverse signaling pathways. Oncogenic mutant K-Ras is the most frequently mutated class of Ras superfamily that is highly prevalent in many human cancers. Despite intensive efforts to combat various K-Ras-mutant-driven cancers, no effective K-Ras-specific inhibitors have yet been approved for clinical use to date. Since K-Ras proteins must be associated to the plasma membrane for their function, targeting K-Ras plasma membrane localization represents a logical and potentially tractable therapeutic approach. Here, we summarize the recent advances in the development of K-Ras plasma membrane localization inhibitors including natural product-based inhibitors achieved from high throughput screening, fragment-based drug design, virtual screening, and drug repurposing as well as hit-to-lead optimizations.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Evaluación Preclínica de Medicamentos , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Proteína Oncogénica p21(ras)/metabolismo , Animales , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/uso terapéutico , Productos Biológicos/química , Productos Biológicos/farmacología , Reposicionamiento de Medicamentos , Humanos , Mutación , Neoplasias/genética , Neoplasias/patología , Proteína Oncogénica p21(ras)/genéticaRESUMEN
We investigated the antitumor effect of Cordyceps militaris extract (CME) on A549 cisplatin-resistant (CR) lung cancer cells. The proliferation of A549/CR cells was suppressed by CME. Apoptosis of the cells was induced by CME. The cell cycle arrest was observed in the sub-G1 phase in the cells treated with CME. Proteomic profile analysis showed that H-Ras was downregulated in CME-treated cells and it was confirmed by western blot analysis. Collectively, these data demonstrated that CME is an alternative treatment for the anticancer effect.
Asunto(s)
Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Cisplatino/farmacología , Cordyceps/química , Neoplasias Pulmonares/fisiopatología , Proteína Oncogénica p21(ras)/genética , Extractos Vegetales/farmacología , Células A549 , Apoptosis/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Resistencia a Antineoplásicos , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Proteína Oncogénica p21(ras)/metabolismoRESUMEN
The achievements in the treatment of metastatic colorectal cancer during recent years are based on a better understanding of the disease and individualized regimen planning. In adjuvant treatment, the highly important IDEA (International Duration Evaluation of Adjuvant Chemotherapy) study has shown that treatment duration can safely be reduced in selected patient populations. In patients with pN1 and pT1-pT3 tumors, 3 months of treatment with 5-fluorouracil and oxaliplatin is comparable with respect to 3-year survival rate to 6 months of treatment. For patients with N2 tumors, 6 months of treatment should stay the standard of care. The limitation of the duration of the adjuvant treatment is significantly reducing the chemotherapy-induced morbidity. New studies will explore the use of immune-checkpoint inhibitors in the adjuvant setting in microsatellite-instable (MSI) tumors. In metastatic disease, next to the required molecular testing for RAS and BRAF mutations, MSI testing is recommended. In the rare group of patients with a MSI tumor, immune-checkpoint inhibition is changing the course of the disease dramatically. Therefore, it is important to identify those patients early. For the RAS-mutant cases, no new and targeted treatment options have been identified yet. An optimal treatment strategy for those patients is urgently needed. RAS wild-type patients with tumors derived from the left side of the colon (splenic flexure to rectum) should be treated in first line with epithelial growth factor receptor (EGFR) antibodies. This selection by a molecular and a clinical marker increased the benefit derived by EGFR antibodies dramatically and defined the most effective treatment option for those patients. New selection criteria based on gene expression, methylation, and other molecular changes are explored and will further influence our therapeutic strategies in the future.
Asunto(s)
Neoplasias Colorrectales/terapia , Anticuerpos/uso terapéutico , Receptores ErbB/inmunología , Fluorouracilo/uso terapéutico , Humanos , Inestabilidad de Microsatélites , Proteína Oncogénica p21(ras)/genética , Oxaliplatino/uso terapéutico , Proteínas Proto-Oncogénicas B-raf/genéticaRESUMEN
Since about 30% of all human cancers contain mutationally activated Ras, down regulating the over-activation of Ras/MAPK pathway represents a viable approach for treating cancers. Over-activation of Ras/MAPK pathway is accompanied by accumulation of reactive oxygen species (ROS). One approach for developing anti-cancer drugs is to target ROS production and their accumulation. To test this idea, we have employed C. elegans of let-60 (gf) mutant, which contain over-activated let-60 (the homolog of mammalian ras) and exhibit tumor-like symptom of multivulva phenotype, to determine whether anti-oxidants can affect their tumor-like phenotype. Specifically we studied the effect of Shengmai formula (SM), a traditional Chinese medicine that has strong anti-oxidant activity, on the physiology of let-60 (gf) mutants. Unexpectedly, we found that SM treatment led to the opening of mitochondrial permeability transition pore by regulating cyclophilin D and then triggered oxidative stress and related signaling pathway activation, including p53, JNK, and p38/MAPK pathways. Finally, SM induced mitochondrial pathway of apoptosis and inhibited the tumor-like symptom of the multivulva phenotype of let-60(gf) mutants. Our results provide evidences to support that SM act as a pro-oxidant agent and could serve as a potential drug candidate for combating over-activated Ras-related cancer.
Asunto(s)
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Ciclofilinas/metabolismo , Medicamentos Herbarios Chinos/farmacología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Animales , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/genética , Peptidil-Prolil Isomerasa F , Ciclofilinas/genética , Combinación de Medicamentos , Sistema de Señalización de MAP Quinasas/genética , Proteínas de Transporte de Membrana Mitocondrial/genética , Poro de Transición de la Permeabilidad Mitocondrial , Mutación , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Neoplasias/metabolismo , Proteína Oncogénica p21(ras)/genética , Proteína Oncogénica p21(ras)/metabolismoRESUMEN
AIMS: Among cancers, hepatocellular carcinoma is one of the commonest worldwide, and its incidence is increasing around the world. A lot of evidence underlines that natural substances usually consumed in the diet can have an important role in the prevention of cancer. In this study we investigated the molecular mechanisms underlying the antiproliferative activity of Citrus bergamia (bergamot) juice (BJ) in human hepatocellular carcinoma HepG2 cells. MAIN METHODS: HepG2 cells were exposed to BJ and then cell proliferation, cell cycle progression, apoptosis and NF-κB nuclear translocation were evaluated. KEY FINDINGS: Here we present results demonstrating that BJ reduced the growth rate of human hepatocellular carcinoma HepG2 cells in a time- and concentration-dependent manner, by a mechanism involving the activation of apoptotic machinery via both intrinsic and extrinsic pathways. Moreover, BJ increased expression of P53 and P21 proteins that may be responsible for the HepG2 cell cycle arrest in G2 phase. In addition, BJ reduced NF-κB nuclear translocation. SIGNIFICANCE: Our data demonstrate the ability of BJ in reducing the growth of HepG2 cells, revealing its mechanism of action and suggesting a promising role as anticancer drugs.
Asunto(s)
Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , FN-kappa B/metabolismo , Extractos Vegetales/farmacología , Ciclo Celular/efectos de los fármacos , Ensayo Cometa , Daño del ADN , ADN Complementario/biosíntesis , ADN Complementario/genética , Fase G2/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/genética , Genes p53/efectos de los fármacos , Células Hep G2 , Compuestos Heterocíclicos/química , Humanos , Proteína Oncogénica p21(ras)/efectos de los fármacos , Proteína Oncogénica p21(ras)/genética , ARN Neoplásico/biosíntesis , Translocación GenéticaRESUMEN
Oncogene-induced senescence (OIS) is characterized by permanent growth arrest and the acquisition of a secretory, pro-inflammatory state. Increasingly, OIS is viewed as an important barrier to tumorgenesis. Surprisingly, relatively little is known about the metabolic changes that accompany and therefore may contribute to OIS. Here, we have performed a metabolomic and bioenergetic analysis of Ras-induced senescence. Profiling approximately 300 different intracellular metabolites reveals that cells that have undergone OIS develop a unique metabolic signature that differs markedly from cells undergoing replicative senescence. A number of lipid metabolites appear uniquely increased in OIS cells, including a marked increase in the level of certain intracellular long chain fatty acids. Functional studies reveal that this alteration in the metabolome reflects substantial changes in overall lipid metabolism. In particular, Ras-induced senescent cells manifest a decline in lipid synthesis and a significant increase in fatty acid oxidation. Increased fatty acid oxidation results in an unexpectedly high rate of basal oxygen consumption in cells that have undergone OIS. Pharmacological or genetic inhibition of carnitine palmitoyltransferase 1, the rate-limiting step in mitochondrial fatty acid oxidation, restores a pre-senescent metabolic rate and, surprisingly, selectively inhibits the secretory, pro-inflammatory state that accompanies OIS. Thus, Ras-induced senescent cells demonstrate profound alterations in their metabolic and bioenergetic profiles, particularly with regards to the levels, synthesis and oxidation of free fatty acids. Furthermore, the inflammatory phenotype that accompanies OIS appears to be related to these underlying changes in cellular metabolism.
Asunto(s)
Senescencia Celular/genética , Metabolismo Energético/genética , Metabolismo de los Lípidos/genética , Oncogenes , Carnitina O-Palmitoiltransferasa/antagonistas & inhibidores , Carnitina O-Palmitoiltransferasa/metabolismo , Línea Celular Tumoral , Proliferación Celular , Citocinas/biosíntesis , Perfilación de la Expresión Génica , Humanos , Inflamación/genética , Metabolómica/métodos , Proteína Oncogénica p21(ras)/genética , Proteína Oncogénica p21(ras)/metabolismo , Consumo de OxígenoRESUMEN
OBJECTIVE: To investigate the effect of total flavonoids of Hedysarum polybotry on the proliferation, cell cycle, and expressions of p21(Ras) and proliferating cell nuclear antigen (PCNA) gene in erythroleukemia cell line K562. METHODS: The effect of total flavonoids of Hedysarum polybotry on K562 cell line survival was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) reduction assay. The time- and dose-dependent manner was also observed. The cell cycle and apoptosis were analyzed with flow cytometry (FCM). The immunocytochemistry method was applied to quantitatively analyze the effects of flavonoids of Hedysarum polybotry on changes p21(Ras) and PCNA gene expressions. RESULTS: Flavonoids of Hedysarum polybotry (20-100 µg/mL) significantly inhibited the proliferation of K562 cells in a time- and dose-dependent manner. After K562 cells were cultured for 48 h, total flavonoids of Hedysarum polybotry had no significant effect on the apoptosis of K562 cells but showed significantly inhibition (P<0.01), indicating that total flavonoids of Hedysarum polybotry could induce K562 cells arrested at G(0)/G(1) and G(2)/M phases. Compared with the control group, p21(Ras) and PCNA gene expressions were decreased significantly in K562 cells treated with total flavonoids of Hedysarum polybotry (40 and 80 µg/mL, respectively) for 48 h. CONCLUSION: The inhibitory effect on proliferation of K562 cells was observed in the groups treated with flavonoids of Hedysarum polybotry, which might be related to cells arresting.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Flavonoides/farmacología , Leucemia Eritroblástica Aguda/tratamiento farmacológico , Proteína Oncogénica p21(ras)/genética , Antígeno Nuclear de Célula en Proliferación/genética , Ranunculaceae/química , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Humanos , Células K562 , Leucemia Eritroblástica Aguda/genética , Leucemia Eritroblástica Aguda/patologíaRESUMEN
Adlay ( Coix lachryma-jobi L. var. ma-yuen Stapf) is a grass crop and was reported to possess anti-inflammatory activity and an antiproliferative effect in cancer cell lines. The purpose of this study was to evaluate the effects of the ethyl acetate fraction of an adlay bran ethanolic extract (ABE-Ea) on colon carcinogenesis in an animal model and investigate its mechanism. Male F344 rats received 1,2-dimethylhydrazine (DMH) and consumed different doses of ABE-Ea. The medium-dose group (17.28 mg of ABE-Ea/day) exhibited the best suppressive effect on colon carcinogenesis and prevented preneoplastic mucin-depleted foci (MDF) formation. Moreover, RAS and Ets2 oncogenes were significantly down-regulated in this group compared to the negative control group, whereas Wee1, a gene involved in the cell cycle, was up-regulated. Cyclooxygenase-2 (COX-2) protein expression was significantly suppressed in all colons receiving the ABE-Ea, indicating that ABE-Ea delayed carcinogenesis by suppressing chronic inflammation. ABE-Ea included considerable a proportion of phenolic compounds, and ferulic acid was the major phenolic acid (5206 microg/g ABE-Ea) on the basis of HPLC analysis. Results from this study suggest that ABE-Ea suppressed DMH-indued preneoplastic lesions of the colon in F344 rats and that ferulic acid may be one of the active compounds.
Asunto(s)
Coix/química , Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/inmunología , Regulación hacia Abajo/efectos de los fármacos , Extractos Vegetales/administración & dosificación , Proteínas Proto-Oncogénicas/genética , 1,2-Dimetilhidrazina/efectos adversos , Acetatos/química , Animales , Antiinflamatorios/administración & dosificación , Neoplasias del Colon/genética , Neoplasias del Colon/patología , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Modelos Animales de Enfermedad , Expresión Génica/efectos de los fármacos , Humanos , Masculino , Proteína Oncogénica p21(ras)/genética , Proteína Oncogénica p21(ras)/inmunología , Proteína Proto-Oncogénica c-ets-2/genética , Proteína Proto-Oncogénica c-ets-2/inmunología , Proteínas Proto-Oncogénicas/inmunología , Ratas , Ratas Endogámicas F344RESUMEN
Epidemiological and experimental carcinogenesis studies provide evidence that certain components of garlic have anti-cancer activity. Although the biotransformed garlic derivative S-allylmercapto-L-cysteine (SAMC) has been reported to show an inhibitory effect on tumorigenesis, the mechanisms are poorly understood. The present study investigated the effect of SAMC on the growth of human gastric cancer SNU-1 cells. Upon treatment with SAMC, a concentration-dependent inhibition of cell proliferation was observed and cells developed many of the hallmark features of apoptosis, including DNA fragmentation and an increase in the sub-diploid population. The anti-proliferative and apoptotic effect of SAMC was associated with the induction of Bax, p53, and caspase-9, rather than the induction of Bcl-2 and p21. Mitochondrial cytochrome c activation and an in vitro caspase-3 activity assay demonstrated that the activation of caspases accompanies the apoptotic effect of SAMC, which mediates cell death. These results suggest that the apoptotic effect of SAMC on gastric cancer SNU-1 cells may be connected with caspase-3 activation through the induction of Bax and p53, rather then Bcl-2 and p21.
Asunto(s)
Apoptosis/efectos de los fármacos , Cisteína/análogos & derivados , Ajo , Allium , Caspasa 3/genética , Caspasa 3/metabolismo , Caspasa 9/genética , Caspasa 9/metabolismo , Caspasas/genética , Caspasas/metabolismo , Ciclo Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cisteína/química , Cisteína/farmacología , Citocromos c/metabolismo , Fragmentación del ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Immunoblotting , Etiquetado Corte-Fin in Situ , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Proteína Oncogénica p21(ras)/genética , Proteína Oncogénica p21(ras)/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismo , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismoRESUMEN
OBJECTIVE: To explore the effect of Buyang Huanwu decoction (BHD) drug serum on rat's in vitro cultured cerebral cortical neuron apoptosis induced by hypoxia, and on the expression of p53 and p21 genes in hypoxia process. METHODS: The model of hypoxia neuron apoptosis was established adopting Daniel method and treated with BHD drug serum. The neuron apoptosis rate was determined by flow cytometry with propidium iodide staining, the p53 and p21 gene expression was tested by immunohistochemical method with flow cytometry. RESULTS: BHD could significantly inhibit the neuron apoptosis induced by hypoxia and down-regulate the expressions of p53 and p21 genes. CONCLUSION: BHD shows inhibition on neuron hypoxia apoptosis and down-regulating of the p53 and p21 gene expression is one of its mechanisms.
Asunto(s)
Corteza Cerebral/patología , Medicamentos Herbarios Chinos/farmacología , Proteína Oncogénica p21(ras)/biosíntesis , Proteína p53 Supresora de Tumor/biosíntesis , Animales , Apoptosis/efectos de los fármacos , Hipoxia de la Célula , Células Cultivadas , Femenino , Masculino , Neuronas/patología , Proteína Oncogénica p21(ras)/genética , Ratas , Ratas Wistar , Suero , Proteína p53 Supresora de Tumor/genéticaRESUMEN
The extract of European mistletoe (Viscum album, L) has been used in adjuvant chemotherapy of cancer and mistletoe lectins are considered to be major active components. The present work was performed to investigate the effects of Korean mistletoe lectin (Viscum album L. coloratum agglutinin, VCA) on proliferation and apoptosis of human hepatoma cells as well as the underlying mechamisms for these effects. We showed that VCA induced apoptosis in both SK-Hep-1 (p53-positive) and Hep 3B (p53-negative) cells through p53- and p21-independent pathways. VCA induced apoptosis by down-regulation of Bcl-2 and by up-regulation of Bax functioning upstream of caspase-3 in both cell lines. In addition, we observed down-regulation of telomerase activity in both VCA-treated cells. Our results provide direct evidence of the anti-tumor potential of this biological response which comes from inhibition of telomerase and consequent inducing apoptosis. VCA-induced apoptosis is regulated by mitochondrial controlled pathway independently of p53. These findings are important for the therapy with preparation of mistletoe because they show that telomerase-dependent mechanism can be targeted by VCA in human hepatocarcinoma. Taken together, our results suggest that the VCA, considered as a telomerase-inhibitor, can be envisaged as a candidate for enhancing sensitivity of conventional anticancer drugs.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/patología , Inhibidores Enzimáticos/farmacología , Genes p53/efectos de los fármacos , Mitocondrias/enzimología , Preparaciones de Plantas , Proteínas de Plantas , Proteínas Proto-Oncogénicas c-bcl-2 , Telomerasa/antagonistas & inhibidores , Toxinas Biológicas/farmacología , Western Blotting , Carcinoma Hepatocelular/enzimología , Caspasa 3 , Inhibidores de Caspasas , Núcleo Celular/patología , Núcleo Celular/ultraestructura , Fragmentación del ADN/efectos de los fármacos , ADN de Neoplasias/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Citometría de Flujo , Genes bcl-2/efectos de los fármacos , Genes bcl-2/genética , Humanos , Corea (Geográfico) , Proteínas de Neoplasias/química , Proteínas de Neoplasias/aislamiento & purificación , Proteína Oncogénica p21(ras)/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Inactivadoras de Ribosomas Tipo 2 , Células Tumorales Cultivadas , Proteína X Asociada a bcl-2RESUMEN
Ras proteins are critical regulators of cell function, including growth, differentiation, and apoptosis, with membrane localization of the protein being a prerequisite for malignant transformation. We have recently demonstrated that feeding fish oil, compared with corn oil, decreases colonic Ras membrane localization and reduces tumor formation in rats injected with a colon carcinogen. Because the biological activity of Ras is regulated by posttranslational lipid attachment and its interaction with stimulatory lipids, we investigated whether docosahexaenoic acid (DHA), found in fish oil, compared with linoleic acid (LA), found in corn oil, alters Ras posttranslational processing, activation, and effector protein function in young adult mouse colon cells overexpressing H-ras (YAMC-ras). We show here that the major n-3 polyunsaturated fatty acid (PUFA) constituent of fish oil, DHA, compared with LA (an n-6 PUFA), reduces Ras localization to the plasma membrane without affecting posttranslational lipidation and lowers GTP binding and downstream p42/44(ERK)-dependent signaling. In view of the central role of oncogenic Ras in the development of colon cancer, the finding that n-3 and n-6 PUFA differentially modulate Ras activation may partly explain why dietary fish oil protects against colon cancer development.
Asunto(s)
Transferasas Alquil y Aril/metabolismo , Transformación Celular Neoplásica , Ácidos Docosahexaenoicos/farmacología , Ácidos Grasos Omega-3/farmacología , Ácidos Grasos Insaturados/farmacología , Genes ras/efectos de los fármacos , Ácido Linoleico/farmacología , Proteína Oncogénica p21(ras)/metabolismo , Animales , División Celular/efectos de los fármacos , División Celular/fisiología , Línea Celular Transformada , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Colon , Aceite de Maíz , Activación Enzimática , Ácidos Grasos Omega-6 , Aceites de Pescado , Lípidos de la Membrana/metabolismo , Ratones , Proteína Oncogénica p21(ras)/genética , Ácido Palmítico/metabolismo , Fosfolípidos/metabolismo , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Ratas , Transducción de Señal/efectos de los fármacosRESUMEN
The chemopreventive effects of tea polyphenols and tea pigments on liver precancerous lesion in rats were investigated. The results showed that the density and area of GST-P in the tea-treated groups were significantly reduced as compared with the positive control group. Furthermore, tea polyphenols and tea pigments induced the expression of p21WAF1 protein, inhibited the expression of Bcl-2 protein and induced the expression of Bax protein. It is concluded that GST-P was significantly inhibited by tea polyphenols and tea pigments and inhibition of cell proliferation and induction of apoptosis may be the two important mechanisms.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Flavonoides , Neoplasias Hepáticas/patología , Fenoles/farmacología , Polímeros/farmacología , Lesiones Precancerosas/patología , Té/química , Animales , Animales Recién Nacidos , Masculino , Proteína Oncogénica p21(ras)/biosíntesis , Proteína Oncogénica p21(ras)/genética , Fenoles/aislamiento & purificación , Pigmentos Biológicos/aislamiento & purificación , Pigmentos Biológicos/farmacología , Polímeros/aislamiento & purificación , Polifenoles , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Proteínas Proto-Oncogénicas c-bcl-2/genética , Ratas , Ratas WistarRESUMEN
OBJECTIVE: To investigate the common regulative effects of the Chinese drug Bailong and hexamethylen bisacetamide (HMBA) on expressions of oncogenes (c-H-ras and c-myc), and tumor suppressor genes (Rb, p53 and p21) of MGC80-3 in human cancer cell cycle. METHODS: Adopting RNA Northern Blot to survey the levels of gene expressions of MGC80-3 different phases cells treated with Bailong and HMBA respectively. RESULTS: In different phases of MGC80-3 cells treated with Bailong and differentiation inducer HMBA, expressions of oncogenes c-H-ras and c-myc were inhibited by over 50.0%, messenger kinase subspecies PKC-alpha gene is similar with the expression inhibition of oncogenes, except effect of Bailong on the G2 phase in cell cycle. Effect of Bailong differs greatly from HMBA in the expression of tumor suppression genes. The expression of Rb and p21 in cells treated by HMBA did not increase but were inhibited by 39.5% and 33.3% respectively in G1 phase. The level of Rb gene expression was decreased, too by 3.0% in S phase. Comparison with HMBA the expression of Rb and p21 genes were increased after treatment by Bailong in all cell cycle. But the effect of Bailong on the expression of p53 gene which was increased obviously by 125.0%-233.4% in majority phase of MGC80-3 cells is similar to HMBA. CONCLUSION: (1) The effect of Bailong on the regulation of oncogenes and tumor suppressor gene is similar to HMBA but the effect of Bailong is better than that of HMBA. (2) Molecular mechanism of the Bailong or HMBA on the proliferative inhibition and differentiation of MGC80-3 related to regulation of the Bailong and HMBA on the oncogenes and tumor suppressor genes in cell cycle of MGC80-3.
Asunto(s)
Acetamidas/farmacología , Medicamentos Herbarios Chinos/farmacología , Genes Supresores de Tumor , Oncogenes , Neoplasias Gástricas/patología , Antineoplásicos/farmacología , Ciclo Celular , Expresión Génica , Humanos , Proteína Oncogénica p21(ras)/biosíntesis , Proteína Oncogénica p21(ras)/genética , Proteínas Proto-Oncogénicas c-myc/biosíntesis , Proteínas Proto-Oncogénicas c-myc/genética , Neoplasias Gástricas/genética , Células Tumorales CultivadasRESUMEN
One major objective of tumor immunologists is to prevent cancer development in individuals at high risk. (TG.AC x C57BL/6)F1 mice serve as a model for testing the feasibility of this objective. The mice carry in the germline a mutant ras oncogene that has an arginine at codon 12 instead of glycine present in the wild-type, and after physical (wounding) or chemical promotion, these mice have a high probability for developing papillomas that progress to cancer. Furthermore, F1 mice immunized with Arg(12) mutant ras peptide in complete Freund's adjuvant (CFA) develop T cells within 10 d that proliferate in vitro on stimulation with the Arg(12) mutant ras peptide. Within 14 d, these mice have delayed-type hypersensitivity to the peptide. Immunization with CFA alone or with a different Arg(12) mutant ras peptide in CFA induced neither response. To determine the effect of immunization on development of tumors, mice immunized 3 wk earlier were painted on the back with phorbol 12-myristate 13-acetate every 3 d for 8 wk. The time of appearance and the number of papillomas were about the same in immunized and control mice, but the tumors grew faster and became much larger in the mice immunized with the Arg(12) mutant ras peptide. Thus, the immunization failed to protect against growth of papillomas. The peptide-induced CD4(+) T cells preferentially recognized the peptide but not the native mutant ras protein. On the other hand, mice immunized with Arg(12) mutant ras peptide and bearing papillomas had serum antibodies that did bind native mutant ras protein. Together, these studies indicate that active immunization of cancer-prone individuals may result in immune responses that fail to eradicate mutant oncogene-expressing tumor cells, but rather induce a remarkable enhancement of tumor growth.
Asunto(s)
Arginina/inmunología , Glicina/inmunología , Proteína Oncogénica p21(ras)/inmunología , Papiloma/inmunología , Mutación Puntual , Animales , Anticuerpos Antineoplásicos/inmunología , Arginina/genética , Vacunas contra el Cáncer/inmunología , Femenino , Glicina/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Proteína Oncogénica p21(ras)/genética , Papiloma/fisiopatología , Péptidos/inmunología , VacunaciónRESUMEN
OBJECTIVE: To study the effect of compound Chinese drug Bailong on the transcription of Cyclin Dependent Kinase Inhibitor (CKI) p16INK4a, p21 and Rb, c-myc genes, and the relationship between gene expression and cAMP-PKA pathway. METHODS: Using the traditional molecular biology methods (cell synchronization, molecular hybridization--Western blotting, Northern blotting, etc.) examine the gene expression. RESULTS: Bailong promoted the expression (both mRNA and protein) of p16INK4a obviously in G1 phase cells. When prekallikrein (PKA) inhibitor was added in the cells which were treated by Bailong, the mRNA and protein level of p16INK4a decreased. It was shown that the inhibited proliferation of BGC82-3 cell by Bailong may come from the enhanced p16INK4a gene expression in G1 phase. Being same as p16INK4a, tumor suppressor genes Rb, p21 and oncogene c-myc expression were all affected by Bailong. When PKA inhibitor was added, the results were reversed. CONCLUSION: Bailong can affect many anticancer genes (including p16INK4a, p21 and Rb genes) and oncogenes (including c-myc) transcription by regulating cAMP-PKA pathway.
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Quinasas Ciclina-Dependientes/antagonistas & inhibidores , Medicamentos Herbarios Chinos/farmacología , Genes Supresores de Tumor , Precalicreína/antagonistas & inhibidores , Transducción de Señal , Neoplasias Gástricas/genética , Genes de Retinoblastoma/genética , Genes myc , Genes p16 , Humanos , Interfase , Proteína Oncogénica p21(ras)/biosíntesis , Proteína Oncogénica p21(ras)/genética , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Células Tumorales CultivadasRESUMEN
Dietary n-6 polyunsaturated fatty acids (PUFAs) promote rat mammary cancer while n-3 PUFAs are inhibitory. The purpose of this study was to determine whether the fats exert their effects by altering the expression of genes that affect cancer development. Therefore, we have examined the effect of PUFAs on the expression of the cyclooxygenase (COX) 1 and 2 genes that are involved in prostaglandin biosynthesis. We also investigated the effect of dietary PUFAs on the expression of the p21ras protein and Ha-ras mRNA. Rats were fed either low- (7%; LF) or high- (21%; HF) fat diets that were rich in either n-6 PUFAs (safflower oil, S) or n-3 PUFAs (menhaden oil, M) for 3 weeks. COX-1 mRNA levels were approximately the same in groups fed diets containing either level of menhaden oil, but were increased by approximately 30% in the LFS and HFS groups (P < 0.05). Transcripts of the inducible COX-2 gene were not detectable in the menhaden oil groups, but this gene was expressed in animals fed either level of safflower oil and in the HFS group was associated with increased levels of COX enzymatic activity and production of PGE2. Animals fed safflower oil had elevated levels of p21ras protein compared to animals fed menhaden oil. Ha-ras mRNA was increased by approximately 35% in animals fed HFS compared to the group fed HFM (P < 0.05). These results demonstrate that dietary n-6 PUFAs upregulate COX-2 and, to some extent, COX-1 expression. There was a concomitant increase in COX enzyme activity and PG synthesis in the mammary glands of rats fed high levels of n-6 PUFAs. Together with associated changes in p21ras expression, these results may explain, at least in part, the promoting effects of dietary n-6 PUFAs on mammary carcinogenesis.
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Grasas de la Dieta/farmacología , Ácidos Grasos Insaturados/farmacología , Isoenzimas/genética , Glándulas Mamarias Animales/efectos de los fármacos , Proteína Oncogénica p21(ras)/genética , Prostaglandina-Endoperóxido Sintasas/genética , Animales , Secuencia de Bases , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Cartilla de ADN , Dinoprostona/metabolismo , Femenino , Genes ras , Glándulas Mamarias Animales/metabolismo , Proteínas de la Membrana , Proteína Oncogénica p21(ras)/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
A ras oncogene-amplified recombinant BHK-21 cell line (ras-rBHK-IgG) has been established, and was shown to hyperproduce the recombinant IgG chimeric human monoclonal antibody (hMAb) AE6F4, which recognizes lung cancer cells. We found that the ras-rBHK-IgG cell could be easily cultured in a protein-free ERDF medium supplemented with iron(III) nitrate, hydroxyethyliminodiacetic acid, and non-protein synthetic attachment factor as well as in a serum-free ERDF medium supplemented with insulin, transferrin, ethanolamine, and sodium selenite. The productivity of recombinant hMAb from the cells cultured in dishes at high cell densities was higher in protein-free medium than in serum-containing medium. True high density culture of the ras-rBHK-IgG cells was done in protein-free medium using the Tecnomouse, which is a novel hollow fiber bioreactor system. After culture for 30 days in protein-free culture, a total amount of about 14 mg of the recombinant hMAb AE6F4 was obtained, and was shown to be reactive against lung cancer cells in tissues.
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Anticuerpos Monoclonales/biosíntesis , Proteína Oncogénica p21(ras)/genética , Animales , Anticuerpos Monoclonales/genética , Células CHO , Línea Celular Transformada , Células Cultivadas , Quelantes/farmacología , Cricetinae , Cricetulus , Medio de Cultivo Libre de Suero , Etanolamina , Etanolaminas/farmacología , Compuestos Férricos/farmacología , Humanos , Iminoácidos/farmacología , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/genética , Insulina/farmacología , Metacrilatos/síntesis química , Metacrilatos/farmacología , Nitratos/farmacología , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Selenito de Sodio/farmacología , Transferrina/farmacologíaRESUMEN
Expression of ras p21 oncoproteins was examined in histological sections of oral squamous cell carcinoma (SCC), epithelial dysplasia, epithelial hyperkeratosis and normal oral mucosa using antibodies to ras p21 with an immunoperoxidase technique. Ras p21-positive staining was found in 47 of 51 (92.2%) cases of oral SCC, 4 of 4 (100%) cases of epithelial dysplasia, 7 of 7 (100%) cases of epithelial hyperkeratosis, and 1 of 6 (16.7%) cases of normal oral mucosa. The positive staining rate of ras p21 in oral SCC, epithelial dysplasia or epithelial hyperkeratosis was significantly higher than that in normal oral mucosa (P < 0.05). No correlation was found between ras p21 expression and patient age, tumour location, tumour size, clinical staging or histological differentiation of SCC. However, a significant positive correlation was found between ras p21 expression and patients' sex (P < 0.05) or regional lymph node status (P < 0.05). A significant positive correlation was also discovered between ras p21 expression and patients' smoking habits (P < 0.01), as well as daily or total betel quid (BQ) consumption (P < 0.05). Of the 47 immunostain-positive SCC patients, specimens from 6 patients were also obtained after chemotherapy, when ras p21 expression was found to be reduced. These results indicate that ras p21 overexpression may play an important role in the initiation and progression of oral SCCs in patients who are smokers and BQ chewers.
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Carcinoma de Células Escamosas/genética , Regulación Neoplásica de la Expresión Génica , Neoplasias de la Boca/genética , Proteína Oncogénica p21(ras)/genética , Lesiones Precancerosas/genética , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Anticuerpos , Areca , Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/patología , Progresión de la Enfermedad , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Técnicas para Inmunoenzimas , Leucoplasia Bucal/genética , Leucoplasia Bucal/patología , Ganglios Linfáticos/patología , Masculino , Persona de Mediana Edad , Mucosa Bucal/metabolismo , Neoplasias de la Boca/tratamiento farmacológico , Neoplasias de la Boca/patología , Estadificación de Neoplasias , Proteína Oncogénica p21(ras)/efectos de los fármacos , Plantas Medicinales , Lesiones Precancerosas/patología , Factores Sexuales , Fumar/genética , Coloración y Etiquetado , TaiwánRESUMEN
Apigenin, a plant flavonoid, induced the reversion of transformed phenotypes of v-H-ras-transformed NIH 3T3 cells at a quite low concentration of 12.5 microM. In the present study, we have examined the components of this Ras-mediated signaling transduction to determine whether they were involved in the apigenin-induced reversion process. Interestingly, the consitutively activated mitogen activated protein kinase (MAPK) in the ras transformant was inhibited significantly and rapidly by 25 microM apigenin within 30 min, and this reduction continued for more than 4 h. Corroborating these observations, expression of the downstream oncogenes c-jun and c-fos was also dramatically reduced during the first 4 h of treatment. We found that the levels of ras protein and mRNA were not affected by 24 h of treatment with apigenin. These findings indicate that apigenin-induced reversion of v-H-ras-transformed NIH 3T3 cells may occur by inhibiting MAPK activity and its downstream oncogenes rather than by affecting the expression of the ras gene.