RESUMO
Breast cancer is the most prevalent neoplasia among women worldwide. For the estrogen receptor-positive (ER+) phenotype, tamoxifen is the standard hormonal therapy; however, it carries the risk of promoting endometrial carcinoma. Hence, we aimed to evaluate the antiproliferative effect of the phytochemical α-mangostin (AM) as a co-adjuvant alongside tamoxifen on breast cancer cells to improve its efficacy while reducing its adverse effects on endometrium. For this, ER+ breast cancer cells (MCF-7 and T-47D) and endometrial cells (N30) were treated with AM, 4-hydroxytamoxifen (4-OH-TMX), and their combination. Cell proliferation was evaluated using sulforhodamine B assay, and the pharmacological interaction was determined through the combination index and the dose reduction index calculation. The genes KCNH1, CCDN1, MKI67, and BIRC5 were amplified by real-time PCR as indicators of oncogenesis, cell cycle progression, cell proliferation, and apoptosis, respectively. Additionally, genes involved in ER signaling were analyzed. In breast cancer cells, the combination of AM with 4-OH-TMX showed a synergistic antiproliferative effect and favorable dose reduction. AM and 4-OH-TMX decreased KCNH1, CCND1, and BIRC5 gene expression. In endometrial cells, AM decreased MKI-67 gene expression, while it reverted the 4-OH-TMX-dependent CCND1 upregulation. This study establishes the benefits of incorporating AM as a co-adjuvant for first-line ER+ breast cancer therapy.
RESUMO
The potentiation of pharmacological effects can be achieved through several strategies, such as the association of substances and delivery in nanostructured systems. In practice, potentiation can be measured by the law of mass action and joint evaluation of the combination index (CI) and dose-response curves. In this context, this study aimed to evaluate the anti-inflammatory effect of the association of ß-caryophyllene and indomethacin in the free form and delivered in nanoemulsions using the in vitro model of LPS-stimulated murine macrophage. The results indicated potentiation of the anti-inflammatory effect of nanoemulsified substances compared to free substances, as well as synergistic action between the sesquiterpene and the selected NSAID. In comparison, the association of ß-caryophyllene and indomethacin in the free form inhibited the production of nitric oxide by 50% at 48.60 µg/mL (CI = 0.21), while the nanoemulsified association of these substances resulted in an IC50 of 1.45 µg/mL (CI = 0.14). In parallel, cytotoxicity assays on HaCaT and MRC-5 cell lines demonstrated the safety of IC50-equivalent concentrations of the anti-inflammatory action, and no irritating effects on the chorioallantoic membrane of embryonated eggs were observed (HET-CAM assay). The results suggest that ß-caryophyllene may be an alternative to replace an inert oily core in nanoemulsion systems when anti-inflammatory effects are desirable.
Assuntos
Indometacina , Lipopolissacarídeos , Camundongos , Animais , Indometacina/farmacologia , Indometacina/metabolismo , Lipopolissacarídeos/farmacologia , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/metabolismo , MacrófagosRESUMO
Abstract Cerebrovascular disease is the second most serious disease in the world. It has the features of high morbidity, high mortality and recurrence rate. Numerous research on the compatibility of Chinese medicine with effective ingredients of cerebral ischemia has been made during the past decades. The purpose of this study is to quantitatively analyze the combined pharmacological effect of effective ingredients in Danshen and Honghua (Dan Hong) on rat microvascular endothelial cells after gradually oxygen-glucose deprivation. The experimental concentration range for the compatibility of two effective ingredients were determined in the preliminary experiments by Cell Counting kit-8 (CCK-8) method. Drugs were added to rat brain microvascular endothelial cells at a non-toxic dose level. After that, the cells were cultured for 12 h, and placed in a hypoxic environment. Finally, the cell survival rate was used as a measure of drug effect. In order to determine synergism or antagonism, the combination index (CI)-isobologram method was performed to analyze the data from the experiments. Based on this theory, the potencies of each drug and the shapes of their does-effect curves are both taken into account. The results show that the synergism or the antagonism between two effective ingredients compatibility change with different proportion and dosage. Furthermore, it can be seen from the results of these experiments that when these drugs are used in combination, the dosage required to achieve the same therapeutic effects is greatly reduced compared with the case of single one. It is worth mentioning that our experiments also prove that the median-effect equation and the CI method can be applied in the field of traditional Chinese medicine.
Assuntos
Animais , Masculino , Feminino , Ratos , Células Endoteliais/classificação , Estudos de Avaliação como Assunto , Preparações Farmacêuticas/administração & dosagem , Transtornos Cerebrovasculares/patologia , Carthamus tinctorius/efeitos adversosRESUMO
Abstract Hypoxia-inducible factors (HIFs) and cancer stem cells (CSCs) are two challenging causes of radiotherapy and chemotherapy resistance, leading to most cases of failure and recurrence in breast cancer therapy. This study was conducted to investigated the inhibitory effect of combination therapy with doxorubicin (an anthracycline) and FM19G11 (an HIF inhibitor) on MCF-7 cells and their CSC-like cells (CSC-LCs). MCF-7 CSC-LCs with a CD44+/CD24- phenotype were sorted and characterized by flow cytometry. A combination of doxorubicin and FM19G11 caused more cytotoxic effects on MCF-7 and CSC-LCs compared to doxorubicin monotherapy. The largest synergistic effect was observed in CSC-LCs under hypoxic conditions; however, MCF-7 cells showed no synergism in normoxic conditions. The administration of doxorubicin and FM19G11 induced late apoptotic and necrotic cell death in MCF-7 and CSC-LCs. Additionally, G2 phase arrest was observed in both cells. Our results demonstrated that co-administration of FM19G11 and doxorubicin had a synergistic effect in hypoxia and improved drug resistance in breast cancer stem cells.
RESUMO
Nutraceutical combinations that act synergistically could be a powerful solution against colon cancer, which is the second deadliest malignancy worldwide. In this study, curcumin (C), sulforaphane (S), and dihydrocaffeic acid (D, a chlorogenic acid metabolite) were evaluated, individually and in different combinations, over the viability of HT-29 and Caco-2 colon cancer cells, and compared against healthy fetal human colon (FHC) cells. The cytotoxic concentrations to kill 50%, 75%, and 90% of the cells (CC50, CC75, and CC90) were obtained, using the MTS assay. Synergistic, additive, and antagonistic effects were determined by using the combination index (CI) method. The 1:1 combination of S and D exerted synergistic effects against HT-29 at 90% cytotoxicity level (doses 90:90 µM), whereas CD(1:4) was synergistic at all cytotoxicity levels (9:36-34:136 µM) and CD(9:2) at 90% (108:24 µM) against Caco-2 cells. SD(1:1) was significantly more cytotoxic for cancer cells than healthy cells, while CD(1:4) and CD(9:2) were similarly or more cytotoxic for healthy cells. Therefore, the SD(1:1) combination was chosen as the best. A model explaining SD(1:1) synergy is proposed. SD(1:1) can be used as a basis to develop advanced food products for the prevention/co-treatment of colon cancer.
Assuntos
Ácidos Cafeicos/farmacologia , Neoplasias do Colo/dietoterapia , Curcumina/farmacologia , Isotiocianatos/farmacologia , Células CACO-2 , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Células HT29 , Humanos , SulfóxidosRESUMO
BACKGROUND: Expression and activity of the potassium channel ether-à-go-go-1 (EAG1) are strongly related to carcinogenesis and tumor progression, which can be exploited for therapeutic purposes. EAG1 activity may be reduced by preventing its phosphorylation with epidermal growth factor receptor (EGFR) kinase inhibitors and by astemizole, which blocks the channel pore and downregulates its gene expression. OBJECTIVE: We aimed to study the potential cooperative antiproliferative effect of the EGFR inhibitor gefitinib and the EAG1-blocker astemizole, in breast cancer cells. MATERIALS AND METHODS: The cells were characterized by immunocytochemistry. Inhibitory concentrations were determined by non-linear regression analysis using dose-response curves. The nature of the pharmacological effect was evaluated by the combination index equation while cell cycle analysis was studied by flow cy-tometry. RESULTS: Astemizole and gefitinib inhibited cell proliferation in a concentration-dependent manner, with inhibitory concentrations (IC 50) values of 1.72 µM and 0.51 µM, respectively. All combinations resulted in a synergistic antiproliferative effect. The combination of astemizole and gefitinib diminished the percentage of cells in G2/M and S phases, while increased accumulation in G0/G1 of the cell cycle. CONCLUSIONS: Astemizole and gefitinib synergistically inhibited proliferation in breast cancer cells expressing both EGFR and EAG1. Our results suggest that the combined treatment increased cell death by targeting the oncogenic activity of EAG1.
Assuntos
Antineoplásicos/farmacologia , Astemizol/farmacologia , Neoplasias da Mama/tratamento farmacológico , Gefitinibe/farmacologia , Antineoplásicos/administração & dosagem , Astemizol/administração & dosagem , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/genética , Canais de Potássio Éter-A-Go-Go/antagonistas & inibidores , Canais de Potássio Éter-A-Go-Go/genética , Feminino , Gefitinibe/administração & dosagem , Regulação Neoplásica da Expressão Gênica , Humanos , Concentração Inibidora 50 , Inibidores de Proteínas Quinases/administração & dosagem , Inibidores de Proteínas Quinases/farmacologiaRESUMO
Abstract Background Expression and activity of the potassium channel ether-à-go-go-1 (EAG1) are strongly related to carcinogenesis and tumor progression, which can be exploited for therapeutic purposes. EAG1 activity may be reduced by preventing its phosphorylation with epidermal growth factor receptor (EGFR) kinase inhibitors and by astemizole, which blocks the channel pore and downregulates its gene expression. Objective We aimed to study the potential cooperative antiproliferative effect of the EGFR inhibitor gefitinib and the EAG1-blocker astemizole, in breast cancer cells. Materials and Methods The cells were characterized by immunocytochemistry. Inhibitory concentrations were determined by non-linear regression analysis using dose-response curves. The nature of the pharmacological effect was evaluated by the combination index equation while cell cycle analysis was studied by flow cytometry. Results Astemizole and gefitinib inhibited cell proliferation in a concentration-dependent manner, with inhibitory concentrations (IC 50) values of 1.72 µM and 0.51 µM, respectively. All combinations resulted in a synergistic antiproliferative effect. The combination of astemizole and gefitinib diminished the percentage of cells in G2/M and S phases, while increased accumulation in G0/G1 of the cell cycle. Conclusions Astemizole and gefitinib synergistically inhibited proliferation in breast cancer cells expressing both EGFR and EAG1. Our results suggest that the combined treatment increased cell death by targeting the oncogenic activity of EAG1.
Assuntos
Humanos , Feminino , Neoplasias da Mama/tratamento farmacológico , Astemizol/farmacologia , Gefitinibe/farmacologia , Antineoplásicos/farmacologia , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Regulação Neoplásica da Expressão Gênica , Astemizol/administração & dosagem , Concentração Inibidora 50 , Linhagem Celular Tumoral , Inibidores de Proteínas Quinases/administração & dosagem , Inibidores de Proteínas Quinases/farmacologia , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Canais de Potássio Éter-A-Go-Go/antagonistas & inibidores , Canais de Potássio Éter-A-Go-Go/genética , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/genética , Gefitinibe/administração & dosagem , Antineoplásicos/administração & dosagemRESUMO
Boll weevil is the major cotton pest in Brazil, and insecticides are widely recommended against it. We determined the susceptibility of boll weevil to insecticides either in single or in mixture ready-to-use formulations, which are registered to spray cotton fields under the hypothesis that mixtures are more toxic to the target pest. Concentration-mortality curves were determined to adult species, simultaneously through dried residues and ingestion. Ten insecticide formulations were studied with five in mixture (lambda-cyhalothrin + thiamethoxam, lambda-cyhalothrin + chlorantraniliprole, thiamethoxam + chlorantraniliprole, and fenitrothion + esfenvalerate) and their five respective single formulations. Cotton leaf discs and cotyledons were dipped into insecticide dilutions prepared by diluting the commercial products into distilled water. Adult mortality was assessed 48 hours after caging adults on treated and untreated materials. The LC50s-concentrations varied from 0.004 to 0.114 g a.i./L, with a relative potency between single and mixture ones, varying from 1.37- to 29.59-fold. Furthermore, lambda-cyhalothrin and thiamethoxam in single formulation were the most toxic insecticides to boll weevil. Among insecticide mixtures, only lambda-cyhalothrin + chlorantraniliprole resulted in a synergic effect; whereas the remaining mixtures showed an antagonistic effect. Therefore, except for the mixture of lambda-cyhalothrin + chlorantraniliprole, the remaining mixtures did not enhance toxicity against the boll weevil and should be recommended only when aimed at different purposes.(AU)
Bicudo-do-algodoeiro é a principal praga do algodoeiro no Brasil, sendo o uso de inseticidas amplamente recomendado para o seu controle. A suscetibilidade do bicudo-do-algodoeiro foi determinada a inseticidas em formulação simples ou em misturas prontas para uso, as quais têm sido recomendadas para pulverizar campos de algodão sob a hipótese de serem mais tóxicas à praga alvo. Assim, curvas de concentração-mortalidade foram determinadas para adultos do bicudo contaminados, simultaneamente, via resíduo seco e ingestão dos inseticidas. Dez formulações foram estudadas, sendo cinco misturas (lambda-cialotrina + tiametoxam, lambda-cialotrina + clorantraniliprole, tiametoxam + clorantraniliprole e fenitrotiona + esfenvalerato) e suas respectivas cinco formulações simples. Folhas e cotilédones do algodoeiro foram mergulhados em diluições do inseticida preparadas com os produtos comerciais e água destilada. A mortalidade adulta foi avaliada 48 horas após o acondicionamento dos adultos em materiais tratados e não tratados. As concentrações de CL50s variaram de 0,004 a 0,114 g i.a./L, com potência relativa entre formulação simples e misturas, variando de 1,37 a 29,59 vezes. A lambda-cialotrina e o tiametoxam em formulações simples foram os inseticidas mais tóxicos para o bicudo. Entre as misturas, aquela preparada com lambda - cialotrina + clorantraniliprole resultou em um efeito sinérgico, enquanto as demais misturas mostraram um efeito antagonista. Portanto, exceto pela mistura de lambda-cialotrina + clorantraniliprole, as demais misturas não demonstraram maior toxicidade para o bicudo-do-algodoeiro e devem ser recomendadas somente quando objetivam finalidades diferentes.(AU)
Assuntos
Pragas da Agricultura , Gossypium , Inseticidas , Controle de PragasRESUMO
Boll weevil is the major cotton pest in Brazil, and insecticides are widely recommended against it. We determined the susceptibility of boll weevil to insecticides either in single or in mixture ready-to-use formulations, which are registered to spray cotton fields under the hypothesis that mixtures are more toxic to the target pest. Concentration-mortality curves were determined to adult species, simultaneously through dried residues and ingestion. Ten insecticide formulations were studied with five in mixture (lambda-cyhalothrin + thiamethoxam, lambda-cyhalothrin + chlorantraniliprole, thiamethoxam + chlorantraniliprole, and fenitrothion + esfenvalerate) and their five respective single formulations. Cotton leaf discs and cotyledons were dipped into insecticide dilutions prepared by diluting the commercial products into distilled water. Adult mortality was assessed 48 hours after caging adults on treated and untreated materials. The LC50s-concentrations varied from 0.004 to 0.114 g a.i./L, with a relative potency between single and mixture ones, varying from 1.37- to 29.59-fold. Furthermore, lambda-cyhalothrin and thiamethoxam in single formulation were the most toxic insecticides to boll weevil. Among insecticide mixtures, only lambda-cyhalothrin + chlorantraniliprole resulted in a synergic effect; whereas the remaining mixtures showed an antagonistic effect. Therefore, except for the mixture of lambda-cyhalothrin + chlorantraniliprole, the remaining mixtures did not enhance toxicity against the boll weevil and should be recommended only when aimed at different purposes.(AU)
Bicudo-do-algodoeiro é a principal praga do algodoeiro no Brasil, sendo o uso de inseticidas amplamente recomendado para o seu controle. A suscetibilidade do bicudo-do-algodoeiro foi determinada a inseticidas em formulação simples ou em misturas prontas para uso, as quais têm sido recomendadas para pulverizar campos de algodão sob a hipótese de serem mais tóxicas à praga alvo. Assim, curvas de concentração-mortalidade foram determinadas para adultos do bicudo contaminados, simultaneamente, via resíduo seco e ingestão dos inseticidas. Dez formulações foram estudadas, sendo cinco misturas (lambda-cialotrina + tiametoxam, lambda-cialotrina + clorantraniliprole, tiametoxam + clorantraniliprole e fenitrotiona + esfenvalerato) e suas respectivas cinco formulações simples. Folhas e cotilédones do algodoeiro foram mergulhados em diluições do inseticida preparadas com os produtos comerciais e água destilada. A mortalidade adulta foi avaliada 48 horas após o acondicionamento dos adultos em materiais tratados e não tratados. As concentrações de CL50s variaram de 0,004 a 0,114 g i.a./L, com potência relativa entre formulação simples e misturas, variando de 1,37 a 29,59 vezes. A lambda-cialotrina e o tiametoxam em formulações simples foram os inseticidas mais tóxicos para o bicudo. Entre as misturas, aquela preparada com lambda - cialotrina + clorantraniliprole resultou em um efeito sinérgico, enquanto as demais misturas mostraram um efeito antagonista. Portanto, exceto pela mistura de lambda-cialotrina + clorantraniliprole, as demais misturas não demonstraram maior toxicidade para o bicudo-do-algodoeiro e devem ser recomendadas somente quando objetivam finalidades diferentes.(AU)
Assuntos
Pragas da Agricultura , Gossypium , Inseticidas , Controle de PragasRESUMO
OBJECTIVE: This work was aimed to investigate the pharmacodynamic interactions between gnaphaliins A and B with ipratropium bromide (IBR) and salbutamol (SAL) using the guinea pig trachea model through application of the combination index (CI)-isobologram equation. METHODS: The guinea pig trachea rings in isolated chamber with Krebs-Henseleit solution (37°C) were contracted with carbachol (3 µm), and then, concentration-relaxant effect curves were constructed for individual drugs and in combination at fixed constant ratios (1 : 1, 3 : 1 and 1 : 3). Median effect and combination index (CI)-isobologram equations were used for determining interactions. KEY FINDINGS: Gnaphaliin A and gnaphaliin B showed clear synergistic interaction with salbutamol, reducing the dose of salbutamol more than sevenfolds to produce the same relaxant effect. However, the combination of either flavonoids with ipratropium bromide showed no interaction. CONCLUSIONS: Applying the combination index-isobologram method, we determined that gnaphaliin A and gnaphaliin B have synergistic effect with salbutamol due probably to their inhibitory effect on phosphodiesterases to maintain high levels of cAMP in the tracheal smooth muscle. However, these compounds did not show any effect with ipratropium.
Assuntos
Albuterol/farmacologia , Broncodilatadores/farmacologia , Flavonoides/farmacologia , Interações Ervas-Drogas , Ipratrópio/farmacologia , Relaxamento Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Traqueia/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Cobaias , Técnicas In Vitro , MasculinoRESUMO
ß-Lapachone is a phytochemotherapeutic originally isolated from Lapacho tree whose extract has been used medicinally for centuries. It is well known that NAD(P)H:quinone oxidoreductase (NQO1) activity is the principal determinant of ß-Lapachone cytotoxicity. As NQO1 is overexpressed in most common carcinomas, recent investigations suggest its potential application against cancer. Photodynamic therapy (PDT) is a clinically approved and rapidly developing cancer treatment. PDT involves the administration of photosensitizer (PS) followed by local illumination with visible light of specific wavelength. In the presence of oxygen molecules, the light illumination of PS can lead to a series of photochemical reactions and consequently the generation of cytotoxic reactive oxygen species (ROS). It has been reported that ß-Lapachone synergistically interacts with ionizing radiation, hyperthermia and cisplatin and that the sensitivity of cells to ß-Lapachone is closely related to the activity of NQO1. So, the present study aimed to investigate the feasibility of PDT to increase the anticancer effect of ß-Lapachone by up-regulating NQO1 expression on breast cancer MCF-7c3 cells. NQO1 expression was evaluated by Western blot analysis at different times after PDT using ME-ALA as PS. The cytotoxicity of the photodynamic treatment and ß-Lapachone alone or in combination was determined by MTT assay and the combination index (CI)-isobologram method and the dose reduction index (DRI) analysis were used to assess the effect of drug combinations. Our studies for the first time demonstrated that the expression of NQO1 is induced 24h after photodynamic treatment. The sensitivity of cancer cells to ß-Lapachone treatment increased 24h after PDT and a synergistic inhibitory effect on MCF-7c3 cells was showed. Taken together, these results lead us to conclude that the synergistic interaction between ß-Lapachone and PDT in killing cells was consistent with the up-regulation of NQO1. The combination of ß-Lapachone and PDT is a potentially promising modality for the treatment of cancer.