RESUMO
Dihydroartemisinin (DHA) has anticancer effects on multiple tumors, including those associated with breast cancer. This study aimed to investigate the mechanism causing DHA-reversing cisplatin (DDP) resistance in breast cancer. Relative mRNA and protein levels were tested using a qRT-PCR and western blot assay. Cell proliferation, viability, and apoptosis were evaluated using colony formation, MTT, and flow cytometry assays, respectively. Interaction of STAT3 and DDA1 was measured via a dual-luciferase reporter assay. The results showed that DDA1 and p-STAT3 levels were dramatically elevated in DDP-resistant cells. DHA treatment repressed proliferation and induced apoptosis of DDP-resistant cells by suppressing STAT3 phosphorylation; the inhibition ability was positively proportional to the DHA concentration. DDA1 knockdown inhibited cyclin expression, promoted G0/G1 phase arrest, restrained cell proliferation, and induced apoptosis of DDP-resistant cells. Furthermore, knockdown of STAT3 restrained proliferation and induced apoptosis and G0/G1 cell cycle arrest of DDP-resistant cells by targeting DDA1. DHA could restrain tumor proliferation of breast cancer via enhancing drug sensitivity of DDP-resistant cells through the STAT3/DDA1 signaling pathway.
Assuntos
Antineoplásicos , Neoplasias da Mama , MicroRNAs , Neoplasias Ovarianas , Feminino , Humanos , Cisplatino/farmacologia , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias Ovarianas/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos/genética , Linhagem Celular Tumoral , Transdução de Sinais/genética , Proliferação de Células , Apoptose/genética , MicroRNAs/metabolismo , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismoRESUMO
Glioblastoma (GBM) is a highly aggressive brain tumor. During screening work, we found a new compound named phragmunis A (PGA), which is derived from the fruitbody of Trogia venenata, exhibits a potential cytotoxic effect on patient-derived recurrent GBM cells and temozolomide (TMZ)-resistant cell lines. The present study was designed to investigate the potential molecular mechanism of the anti-glioma effects of PGA in vitro and in vivo. Studies investigating the mechanism revealed that PGA diminished the binding efficiency of ETS family of transcription factor (ELK1) and Serum response factor (SRF), and suppressed ELK1-SRF complex-dependent transcription, which decreased the transcriptional levels of downstream genes Early growth response protein 1 (EGR1)-Polycomb ring finger (BMI1), thus inducing the imbalanced regulation between Myeloid cell leukaemia-1 (MCL1) and F-Box and WD repeat domain containing 7 (FBXW7). Finally, orthotopic xenograft models were established to confirm the anti-glioma effect of PGA on tumour growth. We showed, for the first time, that the cytotoxic effects of PGA occurred by inducing MCL1 inhibition and FBXW7 activation by blocking ELK1-SRF complex-dependent transcription. The blockage of ELK1-mediated transcription resulted in the suppression of EGR1-BMI1, which led to the upregulation of FBXW7 expression and downregulation of MCL1. These findings suggested that PGA could be a therapeutic drug candidate for the treatment of recurrent GBM by targeting the ELK1-SRF complex.
Assuntos
Proteína 7 com Repetições F-Box-WD/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Extratos Vegetais/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/metabolismo , Linhagem Celular Tumoral , Proteína 7 com Repetições F-Box-WD/metabolismo , Regulação da Expressão Gênica/fisiologia , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Proteína de Sequência 1 de Leucemia de Células Mieloides/efeitos dos fármacos , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Fator de Resposta Sérica/efeitos dos fármacos , Fator de Resposta Sérica/metabolismo , Proteínas Elk-1 do Domínio ets/efeitos dos fármacos , Proteínas Elk-1 do Domínio ets/metabolismoRESUMO
Glioblastoma multiforme (GBM) is the most frequent, lethal, and aggressive tumor of the central nervous system in adults. In this study, we found for the first time that moschamindole (MCD), a rare phenolic amide with 8/6/6/5/5 rings, is a major bioactive constituent derived from Phragmites communis Trin (Poaceae) that exhibits a potential cytotoxic effect on both TMZ-resistant GBM cell lines and xenograft models. MCD-induced intrinsic apoptosis signals and mitochondrial dysfunction were confirmed by cell cycle arrest, caspase-3/7 activation, and membrane potential depolarization. Furthermore, investigations exploring the mechanism showed that MCD specifically inhibits Mia40-mediated oxidative folding of mitochondrial intermembrane space (IMS) proteins via PCR assay and immunoblot analysis. MCD relies on its positive charge to associate with mitochondrial oxidative respiration, thus blocking energy metabolism and inducing apoptosis. Overexpression and upregulation of Mia40 were proven to reverse MCD-induced apoptosis and were correlated with the chemoresistance of GBM in vitro and in vivo, respectively. Taken together, our study demonstrates that Mia40 is a potential target of the chemoresistance of glioblastoma and suggests that MCD might be a potential agent for the individualized treatment of chemoresistant GBM based on mitochondrial metabolic characteristics and Mia40 expression.
Assuntos
Apoptose/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Mitocôndrias/metabolismo , Animais , Glioblastoma/patologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Proteínas do Complexo de Importação de Proteína Precursora Mitocondrial , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: Glioblastoma multiforme (GBM) is the most frequent, lethal and aggressive tumour of the central nervous system in adults. The discovery of novel anti-GBM agents based on the isocitrate dehydrogenase (IDH) mutant phenotypes and classifications have attracted comprehensive attention. PURPOSE: Diterpenoids are a class of naturally occurring 20-carbon isoprenoid compounds, and have previously been shown to possess high cytotoxicity for a variety of human tumours in many scientific reports. In the present study, 31 cassane diterpenoids of four types, namely, butanolide lactone cassane diterpenoids (I) (1-10), tricyclic cassane diterpenoids (II) (11-15), polyoxybutanolide lactone cassane diterpenoids (III) (16-23), and fused furan ring cassane diterpenoids (IV) (24-31), were tested for their anti-glioblastoma activity and mechanism underlying based on IDH1 mutant phenotypes of primary GBM cell cultures and human oligodendroglioma (HOG) cell lines. RESULTS: We confirmed that tricyclic-type (II) and compound 13 (Caesalpin A, CSA) showed the best anti-neoplastic potencies in IDH1 mutant glioma cells compared with the other types and compounds. Furthermore, the structure-relationship analysis indicated that the carbonyl group at C-12 and an α, ß-unsaturated ketone unit fundamentally contributed to enhancing the anti-glioma activity. Studies investigating the mechanism demonstrated that CSA induced oxidative stress via causing glutathione reduction and NOS activation by negatively regulating glutaminase (GLS), which proved to be highly dependent on IDH mutant type glioblastoma. Finally, GLS overexpression reversed the CSA-induced anti-glioma effects in vitro and in vivo, which indicated that the reduction of GLS contributed to the CSA-induced proliferation inhibition and apoptosis in HOG-IDH1-mu cells. CONCLUSION: Therefore, the present results demonstrated that compared with other diterpenoids, tricyclic-type diterpenoids could be a targeted drug candidate for the treatment of secondary IDH1 mutant type glioblastoma through negatively regulating GLS.
Assuntos
Apoptose/efeitos dos fármacos , Neoplasias Encefálicas/patologia , Diterpenos/farmacologia , Glioblastoma/patologia , Glutaminase/antagonistas & inibidores , Isocitrato Desidrogenase/genética , Mutação , Linhagem Celular Tumoral , Humanos , Estresse OxidativoRESUMO
Glioblastoma multiforme (GBM) is a highly aggressive type of brain tumour. Patients with GBM respond poorly to chemotherapy and have poor survival outcomes. Neuron-glial antigen 2 (NG2), also known as chondroitin sulphate proteoglycan 4 (CSPG4), has been shown to contribute to critical processes, such as cell survival, proliferation, and chemotherapy resistance, during glioma progression. In this study, we found that furanodienone (FUR), a diene-type sesquiterpene isolated from the rhizomes of Rhizoma curcumae, exhibited a potential cytotoxic effect on temozolomide (TMZ)-resistant GBM cells in vitro by inhibiting CSPG4 and related signalling pathways. Studies investigating the mechanism demonstrated that FUR suppressed CSPG4-Akt-ERK signalling, inflammatory responses, and cytokine levels but activated caspase-dependent pathways and mitochondrial dysfunction. Furthermore, an immunofluorescence assay and a dual-luciferase reporter assay revealed that inhibition of EGR1-mediated transcription might have contributed to the FUR-dependent blockade of CSPG4 signalling and glioma cell survival. These results established a link between FUR-induced CSPG4 inhibition and the suppression of EGR1-dependent transcription. Attenuation of ERK1/2 and cytokine signalling might have generated the EGR1-dependent negative feedback loop of the CSPG4 pathway during FUR-induced apoptosis. These findings suggested that FUR could be a therapeutic candidate for the treatment of malignant glioma via targeting CSPG4 signalling.
Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Proteína 1 de Resposta de Crescimento Precoce/fisiologia , Furanos/farmacologia , Glioblastoma/tratamento farmacológico , Sesquiterpenos/farmacologia , Temozolomida/uso terapêutico , Adulto , Apoptose/efeitos dos fármacos , Apoptose/genética , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Proteoglicanas de Sulfatos de Condroitina/genética , Proteoglicanas de Sulfatos de Condroitina/metabolismo , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Resistencia a Medicamentos Antineoplásicos/genética , Furanos/administração & dosagem , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioblastoma/genética , Glioblastoma/patologia , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sesquiterpenos/administração & dosagem , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Temozolomida/administração & dosagem , Transcrição Gênica/efeitos dos fármacosRESUMO
A strategy to suppress the expression of the DNA repair enzyme O6methylguanineDNA methyltransferase (MGMT) by inhibition of Wnt/ßcatenin signaling may be useful as a novel treatment for pituitary adenoma. Previous studies have reported that Tanshinone IIA (TSA), a major quinone compound isolated from Salvia miltiorrhiza, had antitumor effects. However, whether TSA has antitumor effects against pituitary adenoma and whether the mechanisms are associated with the Wnt/ßcatenin/MGMT pathway remains to be clarified. In the present study, TSA treatment caused apoptosis in AtT20 cells in a concentrationdependent manner, as demonstrated by cell viability reduction, phophatidylserine externalization detected by Annexin V staining and mitochondrial membrane potential disruption detected by JC1 staining, which were associated with activation of caspase3 and DNA fragmentation detected by TUNEL in AtT20 cells. Tcell factor (TCF)lymphoidenhancing factor (LEF) reporter activity was determined by dual luciferase reporter assay and the interaction between ßcatenin and TCF4 were detected using a coimmunoprecipitation kit. The results indicated TSA treatment increased ßcatenin phosphorylation, inhibited ßcatenin nuclear translocation, reduced ßcatenin/TCF4 complex formation and TCFLEF luciferase reporter activity, and subsequently reduced the expression of cyclin D1 and MGMT. Notably, overexpression of MGMT in ßcatenin knock down AtT20 cells abrogated the TSAmediated effects in AtT20 cells. In conclusion, TSA induced apoptosis via inhibition of Wnt/ßcatenindependent MGMT expression, which may provide novel insights into the understanding of the mechanism of the antitumor effects of Salvia miltiorrhiza.
Assuntos
Abietanos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Corticotrofos/efeitos dos fármacos , Metilases de Modificação do DNA/genética , Enzimas Reparadoras do DNA/genética , Regulação Neoplásica da Expressão Gênica , Salvia miltiorrhiza/química , Proteínas Supressoras de Tumor/genética , beta Catenina/genética , Abietanos/isolamento & purificação , Animais , Antineoplásicos Fitogênicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Corticotrofos/metabolismo , Corticotrofos/patologia , Fragmentação do DNA , Metilases de Modificação do DNA/antagonistas & inibidores , Metilases de Modificação do DNA/metabolismo , Enzimas Reparadoras do DNA/antagonistas & inibidores , Enzimas Reparadoras do DNA/metabolismo , Relação Dose-Resposta a Droga , Humanos , Fator 1 de Ligação ao Facilitador Linfoide/genética , Fator 1 de Ligação ao Facilitador Linfoide/metabolismo , Camundongos , Extratos Vegetais/química , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Fator de Transcrição 4/genética , Fator de Transcrição 4/metabolismo , Proteínas Supressoras de Tumor/antagonistas & inibidores , Proteínas Supressoras de Tumor/metabolismo , Via de Sinalização Wnt/efeitos dos fármacos , beta Catenina/antagonistas & inibidores , beta Catenina/metabolismoRESUMO
OBJECTIVE: To investigate the clinical effect of spanishneedles leaves on middle and severe xerophthalmia of menopausal females. METHOD: This study was a prospective random controlled trial. Ninty-six menopausal females diagnosed with xerophthalmnia (aged from 40 to 50) were randomly divided into in two groups: group A' the spanishneedles leaves group (n=48) and group B' the control group (n=48). Both groups were treated with Forte eye drops. All patients were detected at 3, 7, 28 h before and after treatment to evaluate subjective symptoms, OSDI and four tear film indicators. Variance analysis and differential analysis on sample average or median were made on both groups before and after treatment. RESULT: There were no significant difference in symptom and diction indicators between both groups before treatment. For 28 d after treatment, among middle and severe xerophthalmia samples of the spanishneedles leaves group, the mean differences showed significant improvement compared with that before treatment , OSDI and four tear film indicators also showed improvement to varying degrees. For 28 d after treatment, among middle and severe xerophthalmia samples of the vitamin C group, the mean differences showed no significant improvement compared with that before treatment , OSDI and four tear film indicators also showed no remarkable improvement. There were significant differences in OSDI, BUT, SIT, height of tear meniscus and FL between both groups. CONCLUSION: Spanishneedles leaves can effectively improve symposiums and signs of middle and severe xerophthalmia among menopausal females and thus showing clinical significance to some extent.
Assuntos
Menopausa , Folhas de Planta/química , Plantas Medicinais , Xeroftalmia/tratamento farmacológico , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
OBJECTIVE: To establish nude mice models with the liver metastases of colonic adenocarcinoma and study the effects of Geranium sibirum extracts on them. METHODS: Nude mice liver metastases model of colonic cancer was established with human colonic cancer cells line( Ls 174t) inoculated into mice spleen. 36 nude mice were randomly divided into 3 groups, containing control group, Geranium sibirum extracts group and hydroxycamptothecine group. The weight and size of the mice and growth of the carcinoma were recorded. All specimens were examined histologically. pS2 in blood in nude mice with liver metastasis of colonic carcinoma was detected with nested RT-PCR. RESULTS: The incidence of liver metastasis was 100% in this intrasplenic injection model. The pathological results showed that tumour cells of liver metastases were poorly differentiated human colonic adenocarcinoma. In Geranium sibirum extracts group, the tumor number and the weight of liver metastases were significantly lower than those in hydroxycamptothecine group (P < 0.05). Using semiquantitative examination,in Geranium sibirum extracts group,the relative value of pS2 expression in blood was significantly higher than that in hydroxycamptothecine group (P < 0.05). CONCLUSION: Geranium sibirum extracts can effectively inhibit the occurrence of liver metastases carcinoma and decrease the positive expression of pS2, it also has better effect than hydroxycamptothecine so that Geranium sibirum extracts may become the potential therapeutic strategy for liver metatstases of colonic cancer.
Assuntos
Antineoplásicos/farmacologia , Neoplasias do Colo/patologia , Geranium/química , Neoplasias Hepáticas/prevenção & controle , Extratos Vegetais/farmacologia , Proteínas Supressoras de Tumor/metabolismo , Animais , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Modelos Animais de Doenças , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundário , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Extratos Vegetais/uso terapêutico , Plantas Medicinais/química , Distribuição Aleatória , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Reactive oxygen species (ROS) are increased in inflammatory bowel disease (IBD) and have been implicated as mediators of intestinal inflammation. We investigated the hypothesis that N-acetylcysteine (NAC) as a glutathione (GSH) precursor attenuates disease progression in a murine dextran sodium sulfate (DSS)-induced colitis model. A colitis model was induced by adding 5% DSS into the drinking water for 7 days. BALB/c mice were injiciatur enema with saline, 5-ASA, N-acetylcysteine, respectively, and free drinking water as control group. DSS-treated mice developed severe colitis as shown by bloody diarrhea, weight loss, and pathologic involvement. Colon lengths were significantly decreased in DSS-treated mice with decreased GSH activity too (P < 0.01). ROS in the colon, the level of interleukin 1 beta (IL-1 beta) in colonic mucosa, serum tumor necrosis factor a (TNF-alpha), MPO, and MDA were significantly increased in DSS-treated animals (P < 0.01), with decreased PON1 activity (P < 0.01). However, NAC significantly decreased colonic MPO activity, ROS, TNF-alpha and IL-1 beta levels and increased PON1 activity and GSH concentration. Moreover, NAC attenuated the macroscopic colonic damage and the histopathologic changes-induced by DSS while similar to 5-ASA group. These results suggest that NAC may be effective in the treatment of colitis through its up-regulating PON1 and scavenging oxygen-derived free radicals.
Assuntos
Acetilcisteína/farmacologia , Antioxidantes/farmacologia , Arildialquilfosfatase/metabolismo , Colite/induzido quimicamente , Colite/metabolismo , Sulfato de Dextrana/efeitos adversos , Regulação para Cima/efeitos dos fármacos , Acetilcisteína/uso terapêutico , Animais , Antioxidantes/uso terapêutico , Colite/prevenção & controle , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Glutationa/metabolismo , Interleucina-1beta/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Peroxidase/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE: To observe the effect of volatile oil of amomum (VOA) on the expressions of mastocarcinoma-related peptide (PS2) and platelet activating factor (PAF) in helicobacter pyloriassociated gastritis (HPG) and to analyze its potential mechanism. METHODS: Eighty patients with HPG were randomly assigned to two groups, 42 patients in the treated group treated with 0.5 mL VOA, thrice per day; and the 38 patients in the control group receiving Western tertiary medicinal treatment. Gastroscopic picture and helicobacter pylori (HP) infection (by quick urease and Warthin-Starry stain) of the gastro-membrane, expressions of PS2 and PAF (by immunohistochemical assay and Western blotting) as well as the contents of aminohexose and phospholipid (by Neuhaus method) in the gastric membrane of all patients were detected before treatment and 4 weeks after treatment. The clinical efficacy in the two groups was compared. RESULTS: The total effective rate in the treated group was 88.1%, which was significantly higher than that in the control group (78.9%, P<0.05). After treatment, in the treated group, gastric membranous contents of aminohexose and phospholipid was increased, expression of PS2 elevated but that of PAF lowered, all showing significant difference as compared with those in the control group (P<0.01). In the control group, the expressions of PS2 and PAF changed insignificantly. The radical eliminating rate of HP in the treated group and the control group was insignificantly different between them (76.1% vs. 65.8%, P>0.05). CONCLUSION: The mechanism of VOA for anti-gastritis might be related with its action in increasing the expression of PS2 and decreasing the expression of PAF, and thus regulating the hydrophobicity of the gastric membrane.
Assuntos
Amomum , Mucosa Gástrica/química , Gastrite/tratamento farmacológico , Infecções por Helicobacter/tratamento farmacológico , Helicobacter pylori , Óleos Voláteis/uso terapêutico , Peptídeos/análise , Fator de Ativação de Plaquetas/análise , Adulto , Idoso , Western Blotting , Doença Crônica , Feminino , Gastrite/metabolismo , Infecções por Helicobacter/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Óleos Voláteis/efeitos adversos , Fosfolipídeos/análiseRESUMO
OBJECTIVE: To observe the influences of Jianwei Yuyang (JWYY) granules on expressions of PS2 and platelet activating factor (PAF), and analyze its potential mechanism. METHODS: 76 gastric ulcer patients were final diagnosed by gastroscope. They were randomly devided into 3 groups, including JWYY group (36 cases), Ranitidine group (36 cases) and Normal group (12 cases). Detection on Biospy Specimens of gastric mucosa to obseve the change of PAF, PS2 and the contents of aninohexose and phosphatide in ulcerated gastric mucosa for Immunohischemical and Western blotting. RESULTS: The contents of aninohexose, phosphatide of PU patients increased in JWYY group. There was significant difference between JWYY group and Ranitidine group (P < 0.01). The Immunohischemical and Western blotting methods showed that there was a linear correlation between the the contents of phosphatide, anino-hexose and the expression of PAF, PS2 in PU patients (P < 0.01). CONCLUSION: JWYY granule can prevent the occurrence and relaps of ulcer and affect the hydrophobicity of gastic mucosa and strengthen the stability of myxo-gellayer by reducing the expression of PAF, elevating the expression of PS2 and effecting the contents of phosphatide and aninohexose, that may be one of the mechanisms of JWYY to heal ulcer quickly.