RESUMO
Non-small cell lung cancer (NSCLC), which accounts for approximately 85% of all lung cancer cases, is associated with a poor outcome. Rafoxanide is an anthelmintic drug that inhibits tumor growth in certain malignancies. However, its impact on NSCLC remains unknown. In this study, we examined the effect of rafoxanide on NSCLC and dissected the underlying mechanism. The results showed that rafoxanide significantly inhibited the growth, invasion, and migration of NSCLC cells. Besides, rafoxanide can induce NSCLC cell apoptosis and cell cycle arrest in a dose-dependent manner. RNA-seq analysis revealed that genes associated with endoplasmic reticulum stress (ER) stress responses were activated. Mechanistically, we found Rafoxanide can induce ER stress and activate the unfolded protein response (UPR). Apoptosis was activated by excessive ER stress, and autophagy was activated to partially alleviate ER stress. In vivo, we found that rafoxanide inhibited the growth of A549 and H1299 xenograft mouse models without severe side effects. Collectively, the present study indicates that rafoxanide may be a candidate drug for the treatment of NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Animais , Camundongos , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/patologia , Rafoxanida/farmacologia , Rafoxanida/uso terapêutico , Neoplasias Pulmonares/tratamento farmacológico , Proliferação de Células , Estresse do Retículo Endoplasmático , Apoptose , Linhagem Celular TumoralRESUMO
AIMS: Diffuse large B-cell lymphoma (DLBCL) is one of the most aggressive lymphoid malignancies, which remains incurable, thus warranting the development of new therapies. Our previous study determined that rafoxanide is very effective in treating multiple myeloma (MM). In the present study, we tried to evaluate the effects of rafoxanide on DLBCL, as well as the potential underlying molecular mechanisms. MAIN METHODS: We used CCK-8 assay and flow cytometry to assess cell viability and apoptosis. The proteins and pathways associated with apoptosis and proliferation were evaluated through western blot, and xenograft mice were used as the experimental animal model. We also used the TUNEL assay and immunofluorescence for further analyses. KEY FINDINGS: Treatment with different doses of rafoxanide significantly inhibited cell viability and apoptosis. Additionally, the compound induced cell cycle arrest, reduced mitochondrial membrane potential (Δψm), and stimulated reactive oxygen species (ROS) generation without the influence of normal peripheral blood monocytes (PBMCs). As expected, rafoxanide played a role in regulating these proteins and the PTEN/PI3K/AKT and JNK/c-Jun pathways. Furthermore, immunofluorescence and western blot results showed that rafoxanide upregulated H2AX phosphorylation and then inhibited DNA repair in DLBCL. In the xenograft mouse model, tumor volumes were reduced after intraperitoneal injection with rafoxanide. We also observed that TUNEL positive cells were remarkably increased in rafoxanide-treated tumor tissues. SIGNIFICANCE: These results collectively provide a novel choice to regular treatment for DLBCL patients with poor prognosis.
Assuntos
Antineoplásicos/uso terapêutico , Linfoma Difuso de Grandes Células B/tratamento farmacológico , MAP Quinase Quinase 4/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-jun/metabolismo , Rafoxanida/uso terapêutico , Animais , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Dano ao DNA , Humanos , Linfoma Difuso de Grandes Células B/enzimologia , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/patologia , Masculino , Camundongos , Espécies Reativas de Oxigênio/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Rafoxanide is commonly used as anti-helminthic medicine in veterinary medicine, a main compound of salicylanilide. Previous studies have reported that rafoxanide, as an inhibitor of BRAF V600E mutant protein, inhibits the growth of colorectal cancer, multiple myeloma, and skin cancer. However, its therapeutic effect on gastric cancer (GC) and the potential mechanism has not been investigated. Here, we have found that rafoxanide inhibited the proliferation of GC cells in vitro, arrested the cell cycle in the G0/G1 phase, and promoted apoptosis and autophagy in GC cells. Treatment with specific autophagy inhibitor 3-methyladenine drastically inhibited the apoptotic cell death effect by suppressing the switch from autophagy to apoptosis. Mechanistically, we found that rafoxanide inhibited the growth of GC cells in vitro by inhibiting the activity of the PI3K/Akt/mTOR signaling pathway. This process induced autophagy, which essentially resulted in the apoptosis of GC cells. Results from subcutaneous implanted tumor models in nude mice also indicated that rafoxanide inhibited the growth of GC cells in vivo. Taken together, our findings revealed that rafoxanide inhibited the growth of GC cells both in vitro and vivo, indicating a potential drug candidate for the treatment of GC.
Assuntos
Antineoplásicos/uso terapêutico , Antiplatelmínticos/uso terapêutico , Apoptose , Autofagia , Rafoxanida/uso terapêutico , Transdução de Sinais/efeitos dos fármacos , Neoplasias Gástricas/tratamento farmacológico , Animais , Antineoplásicos/farmacologia , Antiplatelmínticos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Rafoxanida/farmacologia , Serina-Treonina Quinases TOR/metabolismoRESUMO
A novel laboratory anticestode assay was developed using Hymenolepis diminuta in the hamster. The commercial anticestode compounds, praziquantel, bunamidine, and niclosamide were active against patent infections of Hymenolepis diminuta in golden hamsters (Mesocricetus auratus) when given orally at 3.125, 100, and 200 mg/kg, respectively. The gastrointestinal nematode anthelmintics, cambendazole and mebendazole, were active at 50 mg/kg. Rafoxanide (fasciolicide) was active at 25 mg/kg, the lowest level tested. The coccidiostat, nicarbazin, was active at experimental levels (800 mg/kg and up). The anthelmintic-ectoparasiticide (endectocide), ivermectin, was inactive against the tapeworm at 0.5 mg/kg, as expected.
Assuntos
Anticestoides/uso terapêutico , Sistema Digestório/parasitologia , Himenolepíase/tratamento farmacológico , Hymenolepis/efeitos dos fármacos , Administração Oral , Amidinas/administração & dosagem , Amidinas/farmacologia , Amidinas/uso terapêutico , Animais , Anticestoides/administração & dosagem , Anticestoides/farmacologia , Cambendazol/administração & dosagem , Cambendazol/farmacologia , Cambendazol/uso terapêutico , Cricetinae , Modelos Animais de Doenças , Hymenolepis/isolamento & purificação , Insetos Vetores/parasitologia , Intestino Delgado/parasitologia , Ivermectina/administração & dosagem , Ivermectina/farmacologia , Ivermectina/uso terapêutico , Masculino , Mebendazol/administração & dosagem , Mebendazol/farmacologia , Mebendazol/uso terapêutico , Mesocricetus , Nicarbazina/administração & dosagem , Nicarbazina/farmacologia , Nicarbazina/uso terapêutico , Niclosamida/administração & dosagem , Niclosamida/farmacologia , Niclosamida/uso terapêutico , Praziquantel/administração & dosagem , Praziquantel/farmacologia , Praziquantel/uso terapêutico , Rafoxanida/administração & dosagem , Rafoxanida/farmacologia , Rafoxanida/uso terapêutico , Distribuição Aleatória , Tribolium/parasitologiaRESUMO
Female ICR mice, 5 to 6 weeks old, were exposed by stomach tube to 25 metacercarial cysts of Echinostoma caproni per mouse. At 14 days post-exposure, mice were fed by stomach tube clorsulon (1000 mg/kg, 500 mg/kg and 100 mg/ kg) or rafoxanide (50 mg/kg, 25 mg/kg and 5 mg/kg) dissolved in dimethylsulphoxide (DMSO) carrier and mebendazole (1000 mg/kg and 500 mg/ kg) or arprinocid (100 mg/kg and 50 mg/kg) suspended in a 2:1 polyethylene glycol (PEG)/DMSO carrier. All drugs were obtained from Merck Inc. (Rahway, New Jersey, USA) and only single dose regimes were used. Experimentally infected mice that served as controls received either DMSO or 2:1 PEG/DMSO carriers or were not given the carrier. Mice were necropsied 15, 16, 18 and 20 days postexposure to worms. Doses of 100 mg/kg of clorsulon and 50 mg/kg of rafoxanide were 100% effective in eliminating the echinostomes on day 1 post-administration of the anthelmintics. Mebendazole and arprinocid were ineffective in eliminating worms at 1 or 2 days post drug administration.
Assuntos
Anti-Helmínticos/uso terapêutico , Echinostoma/efeitos dos fármacos , Equinostomíase/tratamento farmacológico , Enteropatias Parasitárias/tratamento farmacológico , Adenina/análogos & derivados , Adenina/farmacologia , Adenina/uso terapêutico , Animais , Anti-Helmínticos/farmacologia , Dimetil Sulfóxido , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Portadores de Fármacos , Feminino , Intestino Delgado/parasitologia , Mebendazol/farmacologia , Mebendazol/uso terapêutico , Camundongos , Camundongos Endogâmicos ICR , Polietilenoglicóis , Rafoxanida/farmacologia , Rafoxanida/uso terapêutico , Sulfanilamidas/farmacologia , Sulfanilamidas/uso terapêuticoRESUMO
Female ICR mice, 5 to 6 weeks old, were exposed by stomach tube to 25 metacercarial cysts of Echinostoma caproni per mouse. At 14 days post-exposure, mice were fed by stomach tube clorsulon (1000 mg/kg, 500 mg/kg and 100 mg/kg) or rafoxanide (50 mg/kg, 25 mg/kg and 5 mg/kg) dissolved in dimethylsulphoxide (DMSO) carrier and mebendazole (1000 mg/kg and 500 mg/kg) or arprinocid (100 mg/kg and 50 mg/kg) suspended in a 2:1 polyethylene glycol (PEG)/DMSO carrier. All drugs were obtained from Merck Inc. (Rahway, New Jersey, USA) and only single dose regimes were used. Experimentally infected mice that served as controls received either DMSO or 2:1 PEG/DMSO carriers or were not given the carrier. Mice were necropsied 15v, 16, 18 and 20 days postexposure to worms. Doses of 100 mg/kg of clorsulon and 50 mg/kg of rafoxanide were 100% effective in eliminating the echinostomes on day 1 post-administration of the anthelmintics. Mebendazole and arprinocid were ineffective in eliminating worms at 1 or 2 days post drug administration.
Assuntos
Adenina/análogos & derivados , Anti-Helmínticos/uso terapêutico , Equinostomíase/tratamento farmacológico , Mebendazol/uso terapêutico , Rafoxanida/uso terapêutico , Sulfanilamidas/uso terapêutico , Adenina/uso terapêutico , Animais , Relação Dose-Resposta a Droga , Feminino , Camundongos , Camundongos Endogâmicos ICRRESUMO
An enzyme-linked immunosorbent assay (ELISA) was developed using microtiter plates for the immunodiagnosis of fascioliasis in rats and rabbits using extracts of adult worms partially purified by gel filtration chromatography using Sephacryl S-200. Partial purification was necessary to eliminate cross-reactivity with antisera having antibodies to schistosomes. Soft polyvinyl plates clearly gave superior results over hard polystyrine plates. Titers rose by 4 weeks of infection in rats with fascioliasis, by 6 weeks in the case of rabbits, and remained high through at least 12 and 28 weeks, respectively. Titers drop rapidly when animals are successfully treated with a fasciolicidal drug at 4--6 weeks of infection. The results show that the ELISA can be employed for the serodiagnosis of fascioliasis in rats and rabbits and is useful for the prediction of chemotherapeutic success.
Assuntos
Ensaio de Imunoadsorção Enzimática , Fasciolíase/diagnóstico , Técnicas Imunoenzimáticas , Animais , Anticorpos/análise , Fasciola hepatica/imunologia , Fasciolíase/tratamento farmacológico , Fasciolíase/imunologia , Coelhos , Rafoxanida/uso terapêutico , RatosRESUMO
Sheep infected with adult Fasciola hepatica were drenched with rafoxanide. At 4, 8, 16 and 24 h after drenching the sheep were killed and the flukes removed, washed and rapidly frozen in liquid nitrogen. The content of key metabolites in the fermentation pathway were determined and compared with those in control F. hepatica, whose hosts were not treated with rafoxanide. Rafoxanide decreased glycogen, malate, NADH and ATP levels. The level of other metabolites in the pathway increased for the first 8-16 h after rafoxanide treatment. The marked decrease in ATP and glycogen, and the increase in total [NAD+]/[NADH] and [oxaloacetate]/[malate], together with the changed content of other metabolites, led to the conclusion that the mode of action of rafoxanide against F. hepatica in vivo is by uncoupling oxidative phosphorylation.