RESUMO
Melanoma is the most aggressive among all types of skin cancers. The current strategies against melanoma utilize BRAFV600E, as a focal point for targeted therapy. However, therapy resistance developed in melanoma patients against the conventional anti-melanoma drugs hinders the ultimate benefits of targeted therapies. A major mechanism by which melanoma cells attain therapy resistance is via the activation of microphthalmia-associated transcription factor-M (MITF-M), the key transcription factor and oncogene aiding the survival of melanoma cells. We demonstrate that tryptanthrin (Tpn), an indole quinazoline alkaloid, which we isolated and characterized from Wrightia tinctoria, exhibits remarkable anti-tumor activity towards human melanoma through the down-regulation of MITF-M. Microarray analysis of Tpn-treated melanoma cells followed by a STRING protein association network analysis revealed that differential expression of genes in melanoma converges at MITF-M. Furthermore, in vitro and in vivo studies conducted using melanoma cells with differential MITF-M expression status, endogenously or ectopically, demonstrated that the anti-melanoma activity of Tpn is decisively contingent on its efficacy in down-regulating MITF-M expression. Tpn potentiates the degradation of MITF-M via the modulation of MEK1/2-ERK1/2-MITF-M signaling cascades. Murine models demonstrate the efficacy of Tpn in attenuating the migration and metastasis of melanoma cells, while remaining pharmacologically safe. In addition, Tpn suppresses the expression of mutated BRAFV600E and inhibits Casein Kinase 2α, a pro-survival enzyme that regulates ERK1/2 homeostasis in many tumor types, including melanoma. Together, we point to a promising anti-melanoma drug in Tpn, by virtue of its attributes to impede melanoma invasion and metastasis by attenuating MITF-M.
Assuntos
Melanoma , Fator de Transcrição Associado à Microftalmia , Animais , Linhagem Celular Tumoral , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Humanos , Sistema de Sinalização das MAP Quinases , Melanoma/genética , Camundongos , Fator de Transcrição Associado à Microftalmia/genética , Fator de Transcrição Associado à Microftalmia/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Proteínas Proto-Oncogênicas B-raf/genética , Proteínas Proto-Oncogênicas B-raf/metabolismo , QuinazolinasRESUMO
Interferon regulated genes (IRGs) are critical in controlling virus infections. Here, we analyzed the expression profile of IRGs in the brain tissue in a mouse model of chikungunya virus (CHIKV) neurovirulence. Neurovirulence is one of the newer complications identified in disease caused by re-emerging strains of CHIKV, an alphavirus with positive-strand RNA in the Togaviridae family. In microarray analysis, we identified significant upregulation of 269 genes, out of which a predominant percentage (76%) was IRGs. The highly modulated IRGs included Ifit1, Ifi44, Ddx60, Usp18, Stat1, Rtp4, Mnda, Gbp3, Gbp4, Gbp7, Oasl2, Oas1g, Ly6a, Igtp, and Gbp10, along with many others exhibiting lesser changes in expression levels. We found that these IRG mRNA transcripts are modulated in parallel across CHIKV-infected mouse brain tissues, human neuronal cell line IMR-32 and hepatic cell line Huh-7. The genes identified to be highly modulated both in mouse brain and human neuronal cells were Ifit1, Ifi44, Ddx60, Usp18, and Mnda. In Huh-7 cells, however, only two IRGs (Gbp4 and Gbp7) showed a similar level of upregulation. Concordant modulation of IRGs in both mice and human cells indicates that they might play important roles in regulating CHIKV replication in the central nervous system (CNS). The induction of several IRGs in CNS during infection underscores the robustness of IRG-mediated innate immune response in CHIKV restriction. Further studies on these IRGs would help in evolving possibilities for their targeting in host-directed therapeutic interventions against CHIKV.
Assuntos
Febre de Chikungunya/genética , Vírus Chikungunya/imunologia , Interações Hospedeiro-Patógeno , Fatores Reguladores de Interferon/genética , RNA Mensageiro/genética , Transcriptoma/imunologia , Proteínas Virais/genética , Proteínas Adaptadoras de Transdução de Sinal , Animais , Antígenos/genética , Antígenos/imunologia , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos de Diferenciação Mielomonocítica/imunologia , Encéfalo/imunologia , Encéfalo/virologia , Proteínas de Transporte/genética , Proteínas de Transporte/imunologia , Linhagem Celular , Febre de Chikungunya/imunologia , Febre de Chikungunya/virologia , Vírus Chikungunya/patogenicidade , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/imunologia , RNA Helicases DEAD-box/genética , RNA Helicases DEAD-box/imunologia , Modelos Animais de Doenças , Endopeptidases/genética , Endopeptidases/imunologia , Regulação da Expressão Gênica , Hepatócitos/imunologia , Hepatócitos/virologia , Humanos , Imunidade Inata , Fatores Reguladores de Interferon/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Análise em Microsséries , Neurônios/imunologia , Neurônios/virologia , RNA Mensageiro/imunologia , Proteínas de Ligação a RNA , Transdução de Sinais , Fatores de Transcrição/genética , Fatores de Transcrição/imunologia , Ubiquitina Tiolesterase , Proteínas Virais/imunologiaRESUMO
BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the world with an alarming rate of mortality. Despite the advancement in treatment strategies and drug developments, the overall survival rate remains poor. Therefore, it is imperative to develop alternative or complimentary anti cancer drugs with minimum off target effects. Urolithin A, a microbial metabolite of ellagic acid and ellagitannins produced endogenously by human gut micro biome is considered to have anti-cancerous activity. However anti tumorigenic effect of urolithin A in OSCC is yet to be elucidated. In this study, we examined whether urolithin A inhibits cell growth and induces both apoptosis and autophagy dependent cell death in OSCC cell lines. PURPOSE: The present study aims to evaluate the potential of urolithin A to inhibit OSCC and its regulatory effect on OSCC proliferation and invasion in vitro and in vivo mouse models. METHODS: We evaluated whether urolithin A could induce cell death in OSCC in vitro and in vivo mouse models. RESULTS: Flow cytometric and immunoblot analysis on Urolithin A treated OSCC cell lines revealed that urolithin A markedly induced cell death of OSCC via the induction of endoplasmic reticulum stress and subsequent inhibition of AKT and mTOR signaling as evidenced by decreased levels of phosphorylated mTOR and 4EBP1. This further revealed a possible cross talk between apoptotic and autophagic signaling pathways. In vivo study demonstrated that urolithin A treatment reduced tumor size and showed a decrease in mTOR, ERK1/2 and Akt levels along with a decrease in proliferation marker, Ki67. Taken together, in vitro as well as our in vivo data indicates that urolithin A is a potential anticancer agent and the inhibition of AKT/mTOR/ERK signalling is crucial in Urolithin A induced growth suppression in oral cancer. CONCLUSION: Urolithin A exerts its anti tumorigenic activity through the induction of apoptotic and autophagy pathways in OSCC. Our findings suggest that urolithin A markedly induced cell death of oral squamous cell carcinoma via the induction of endoplasmic reticulum stress and subsequent inhibition of AKT and mTOR signaling as evidenced by decreased levels of phosphorylated mTOR and 4EBP1. Urolithin A remarkably suppressed tumor growth in both in vitro and in vivo mouse models signifying its potential as an anticancer agent in the prevention and treatment of OSCC. Henceforth, our findings provide a new insight into the therapeutic potential of urolithin A in the prevention and treatment of OSCC.
Assuntos
Apoptose , Autofagia , Cumarínicos , Neoplasias Bucais , Proteínas Proto-Oncogênicas c-akt , Serina-Treonina Quinases TOR , Animais , Cumarínicos/farmacologia , Humanos , Apoptose/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Autofagia/efeitos dos fármacos , Neoplasias Bucais/tratamento farmacológico , Linhagem Celular Tumoral , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/metabolismo , Camundongos , Camundongos Nus , Camundongos Endogâmicos BALB C , Proliferação de Células/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Antineoplásicos Fitogênicos/farmacologiaRESUMO
Water resource management relies heavily on reliable water quality predictions. Predicting water quality metrics in the watershed system, including dissolved oxygen (DO), is the main emphasis of this work. The enhanced long short-term memory (LSTM) model was suggested to improve the model's performance. Additionally, a hybrid model was employed to calculate the ideal parameter values for the LSTM model, which helped overcome the nonstationarity, unpredictability, and nonlinearity of the data about the water quality parameters. This model recruited the COOT method. The original weekly water quality values at the Vaigai River, Madurai, Tamil Nadu, India, were tested using the suggested hybrid model. An independent LSTM, the hybrid optimisation method takes its cues from the cuckoo bird's reproductive strategy and a novel meta-heuristic optimisation technique dubbed COOT, which is based on the behaviour of a flock of coot birds. If implemented, the suggested hybrid model might serve as an alternate framework for water quality prediction, laying the groundwork for basin-wide efforts to manage water quality and control pollutants.
RESUMO
Polyp detection is a challenging task in the diagnosis of Colorectal Cancer (CRC), and it demands clinical expertise due to the diverse nature of polyps. The recent years have witnessed the development of automated polyp detection systems to assist the experts in early diagnosis, considerably reducing the time consumption and diagnostic errors. In automated CRC diagnosis, polyp segmentation is an important step which is carried out with deep learning segmentation models. Recently, Vision Transformers (ViT) are slowly replacing these models due to their ability to capture long range dependencies among image patches. However, the existing ViTs for polyp do not harness the inherent self-attention abilities and incorporate complex attention mechanisms. This paper presents Polyp-Vision Transformer (Polyp-ViT), a novel Transformer model based on the conventional Transformer architecture, which is enhanced with adaptive mechanisms for feature extraction and positional embedding. Polyp-ViT is tested on the Kvasir-seg and CVC-Clinic DB Datasets achieving segmentation accuracies of 0.9891 ± 0.01 and 0.9875 ± 0.71 respectively, outperforming state-of-the-art models. Polyp-ViT is a prospective tool for polyp segmentation which can be adapted to other medical image segmentation tasks as well due to its ability to generalize well.
Assuntos
Pólipos , Humanos , Instituições de Assistência Ambulatorial , Erros de Diagnóstico , Fontes de Energia Elétrica , Colo , Processamento de Imagem Assistida por ComputadorRESUMO
INTRODUCTION: Sorafenib (Sor) is the first-line treatment option in clinics for treating advanced unresectable hepatocellular carcinoma (HCC). However, acquired chemoresistance and adverse side effects associated with Sor monotherapy limit its clinical benefits. We have previously reported the exceptional anti-HCC potential of uttroside B (Utt-B), a furostanol saponin isolated in our lab from Solanum nigrum Linn. leaves. The current study has evaluated the supremacy of a combinatorial regimen of Sor and Utt-B over Sor monotherapy. METHODS: MTT assay was used for In vitro cytotoxicity studies. A clonogenic assay was conducted to assess the anti-proliferative effect of the combination. Annexin V/PI staining, confocal microscopy, FACS cell cycle analysis, and Western blotting experiments were performed to validate the pro-apoptotic potential of the combination in HepG2 and Huh7 cell lines. Pharmacological safety evaluation was performed in Swiss albino mice. RESULTS: Our results indicate that Utt-B augments Sor-induced cytotoxicity in HepG2 and Huh7 cells. The combination inhibits the proliferation of liver cancer cells by inducing apoptosis through activation of the caspases 7 and 3, leading to PARP cleavage. Furthermore, the combination does not induce any acute toxicity in vivo, even at a dose five times that of the effective therapeutic dose. CONCLUSION: Our results highlight the potential of Utt-B as an effective chemosensitizer, which can augment the efficacy of Sor against HCC and circumvent Sor-induced toxic side effects. Moreover, this is the first and only report to date on the chemosensitizing potential of Utt-B and the only report that demonstrates the therapeutic efficacy and pharmacological safety of a novel combinatorial regimen involving Utt-B and Sor for combating HCC.
RESUMO
[This corrects the article DOI: 10.3389/fonc.2021.656804.].
RESUMO
Lipomas are benign mesenchymal tumors of mature adipocytes and commonly occur in the upper trunk. Mesenteric lipomas are relatively rare tumors of the gastrointestinal system. They are usually asymptomatic, detected incidentally on abdominal imaging, or can present with variable symptoms depending on the location, size, and rapidity of tumor growth. The ileal mesentery is the most common site, occurring in adults ranging from 40 to 60 years. We present the case of an unusually large mesenteric lipoma in an adult male patient who presented with vague abdominal pain.
RESUMO
BACKGROUND: Cystic Hypersecretory Carcinoma (CHC) is a rare subset of breast carcinoma. It is part of a spectrum of cystic hypersecretory lesions which includes cystic hypersecretory hyperplasia (CHH), CHH with atypia, CHC in situ and CHC with invasion. Approximately 65 cases of cystic hypersecretory lesions have been reported; most of them were CHC in situ and only 19 cases of CHC with invasion have been reported so far. CASE PRESENTATION: We are reporting 2 cases of 47 and 62 year old women with a palpable breast mass for 6 and 1 month duration respectively. Trucut biopsy was carried out for both which showed high grade ductal carcinoma in situ with microinvasion in the first patient and the latter showed a tiny focus of invasive carcinoma. Simple mastectomy and modified radical mastectomy (MRM) were done for the respective cases; both showed dilated cystic spaces filled with eosinophilic secretions (thyroid colloid-like), lining neoplastic cells that showed variable degrees of proliferation, atypia and in situ carcinoma. There were foci of invasion in both cases and hence a morphological diagnosis of CHC with invasion was made. CONCLUSION: Owing to a smaller number of reported cases, little is known about the biological behavior, prognosis and molecular profile of cystic hypersecretory carcinoma.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/secundário , Mama/patologia , Carcinoma/diagnóstico por imagem , Carcinoma/secundário , Biópsia , Mama/diagnóstico por imagem , Neoplasias da Mama/classificação , Neoplasias da Mama/cirurgia , Carcinoma in Situ/patologia , Feminino , Técnicas Histológicas , Humanos , Mastectomia , Pessoa de Meia-IdadeRESUMO
The presentation of the COVID19 has endangered several million lives worldwide causing thousands of deaths every day. Evolution of COVID19 as a pandemic calls for automated solutions for initial screening and treatment management. In addition to the thermal scanning mechanisms, findings from chest X-ray imaging examinations are reliable predictors in COVID19 detection, long-term monitoring and severity evaluation. This paper presents a novel deep transfer learning based framework for COVID19 detection and segmentation of infections from chest X-ray images. It is realized as a two-stage cascaded framework with classifier and segmentation subnetwork models. The classifier is modeled as a fine-tuned residual SqueezeNet network, and the segmentation network is implemented as a fine-tuned SegNet semantic segmentation network. The segmentation task is enhanced with a bioinspired Gaussian Mixture Model-based super pixel segmentation. This framework is trained and tested with two public datasets for binary and multiclass classifications and infection segmentation. It achieves accuracies of 99.69% and 99.48% for binary and three class classifications, and a mean accuracy of 83.437% for segmentation. Experimental results and comparative evaluations demonstrate the superiority of this unified model and signify potential extensions for biomarker definition and severity quantization.
RESUMO
A 35-year-old male presented with fatigue for 1 month and was found to have megaloblastic anaemia. Further evaluation showed low globulin levels and pan hypogammaglobulinemia. Past history was significant for chronic small bowel diarrhoea and bilateral genu valgum deformity from childhood. Hence, a malabsorption syndrome with a probable antibody deficiency was suspected. An upper gastrointestinal (GI) endoscopy was done, which revealed chronic atrophic gastritis with Helicobacter pylori infection, dysplasia and subtotal villous atrophy with a paucity of plasma cells, which was suggestive of common variable immunodeficiency (CVID)-related enteropathy. CVID can present with predominantly autoimmune GI manifestations without any history of recurrent infections. The risk of gastric dysplasia and malignancy is high in CVID and needs close monitoring.
Assuntos
Imunodeficiência de Variável Comum , Globulinas , Infecções por Helicobacter , Helicobacter pylori , Médicos , Adulto , Criança , Imunodeficiência de Variável Comum/complicações , Imunodeficiência de Variável Comum/diagnóstico , Infecções por Helicobacter/patologia , Humanos , MasculinoRESUMO
[This corrects the article DOI: 10.3389/fonc.2022.903832.].
RESUMO
The ethnomedicinal plant from the Cucurbitaceae family, Corallocarpus epigaeus, or its bioactive derivatives have been widely utilized in traditional medicine owing to their distinct applications against various human ailments and have lured the interest of ethnobotanists and biochemists. Here, we report for the first time, the anti-cancer potential of a bio-active fraction isolated from the dried rhizome of C. epigaeus, and the bioactive principle identified as cucurbitacin B (Cu-B). The purification processes involving the utilization of multiple organic extracts of C. epigaeus rhizome powder, yielded Cu-B from the Ethyl acetate Cytotoxic Fraction (ECF), obtained by the chromatographic separation of the ethyl acetate extract. Amongst the various cancer lines tested, melanoma cells exhibit maximal sensitivity towards the Cu-B-containing ECF fraction. Cu-B induces an apoptotic mode of cell death initiated intrinsically as well as extrinsically in A375 melanoma cells whilst remaining comparatively less toxic to normal skin fibroblasts. In vivo studies involving a NOD-SCID murine model of human melanoma demonstrate the ability of Cu-B to attenuate tumor growth, while being pharmacologically safe in vivo, as assessed in Swiss albino mice. Furthermore, Cu-B inhibits MEK 1/2 as well as the constitutive and EGF-induced ERK 1/2 activation, indicating a definitive involvement of MAPK signal transducers in regulating Cu-B-mediated anti-melanoma activity. Together, our study demonstrates the anti-melanoma potential of C. epigaeus-derived Cu-B, which indicates the Cucurbitaceae succulent as a prospective source for deriving potent and pharmacologically safe anti-cancer compounds.
RESUMO
Our previous study has demonstrated that Uttroside B (Utt-B), a saponin isolated from the leaves of Solanum nigrum Linn induces apoptosis in hepatic cancer cells and exhibits a remarkable growth inhibition of Hepatocellular Carcinoma (HCC). Our innovation has been granted a patent from the US (US 2019/0160088A1), Canada (3,026,426.), Japan (JP2019520425) and South Korea (KR1020190008323) and the technology have been transferred commercially to Q Biomed, a leading US-based Biotech company. Recently, the compound received approval as 'Orphan Drug' against HCC from US FDA, which reveals the clinical relevance of evaluating its antitumor efficacy against HCC. In the present study, we report that Utt-B promotes pro-survival autophagy in hepatic cancer cells as evidenced by the increased expression of autophagy-related proteins, including LC3-II, Beclin1, ATG 5, and ATG 7, as well as a rise in the autophagic flux. Hence, we investigated whether Utt-B-induced autophagic response is complementing or contradicting its apoptotic program in HCC. Inhibition of autophagy using the pharmacological inhibitors, Bafilomycin A1(Baf A1), and 3-methyl adenine (3-MA), and the biological inhibitor, Beclin1 siRNA, significantly enhances the apoptosis of hepatic cancer cells and hence the cytotoxicity induced by Utt-B. We also found increased expression of autophagy markers in Utt-B-treated xenografts derived from HCC. We further analyzed whether the antimalarial drug, Chloroquine (Cqn), a well-known autophagy inhibitor, can enhance the anticancer effect of Utt-B against HCC. We found that inhibition of autophagy using Cqn significantly enhances the antitumor efficacy of Utt-B in vitro and in vivo, in NOD SCID mice bearing HCC xenografts. Taken together, our results suggest that the antitumor effect of Utt-B against HCC can be further enhanced by blocking autophagy. Furthermore, Utt-B in combination with Cqn, a clinically approved drug, if repurposed and used in a combinatorial regimen with Utt-B, can further improve the therapeutic efficacy of Utt-B against HCC.
RESUMO
[This corrects the article DOI: 10.3389/fonc.2022.812598.].
RESUMO
We previously reported the remarkable potency of uttroside B (Utt-B), saponin-isolated and characterized in our lab from Solanum nigrum Linn, against HCC. Recently, the U.S. FDA approved Utt-B as an 'orphan drug' against HCC. The current study validates the superior anti-HCC efficacy of Utt-B over sorafenib, the first-line treatment option against HCC. The therapeutic efficacies of Utt-B vs. sorafenib against HCC were compared in vitro, using various liver cancer cell lines and in vivo, utilizing NOD.CB17-Prkdcscid/J mice bearing human HCC xenografts. Our data indicate that Utt-B holds an augmented anti-HCC efficacy over sorafenib. Our previous report demonstrated the pharmacological safety of Utt-B in Chang Liver, the normal immortalized hepatocytes, and in the acute and chronic toxicity murine models even at elevated Utt-B concentrations. Here, we show that higher concentrations of sorafenib induce severe toxicity, in Chang Liver, as well as in acute and chronic in vivo models, indicating that, apart from the superior therapeutic benefit over sorafenib, Utt-B is a pharmacologically safer molecule, and the drug-induced undesirable effects can, thus, be substantially alleviated in the context of HCC chemotherapy. Clinical studies in HCC patients utilizing Utt-B, is a contiguous key step to promote this drug to the clinic.
RESUMO
BACKGROUND: Tinospora cordifolia (Willd).Miers is a perennial climbing medicinal shrub that has been traditionally used for the treatment of chronic inflammatory ailments. Our previous pre- clinical studies on anti-inflammatory effects, proved that the chloroform extract of T. cordifolia (CETC) suppressed the LPS induced up-regulation of pro-inflammatory biomarkers, hence, further follow up study was carried out to evaluate whether CETC can exhibit a protective effect against LPS induced lethal endotoxemia in vivo and also to analyze the impact of CETC pre-treatment on the secretion of pro-inflammatory cytokines in vitro by THP-1 cells. METHODS: To corroborate our previous preclinical studies on inflammation, we investigated the mechanism of the anti-inflammatory effect of T. cordifolia on THP-cells which were pre-incubated with CETC (30 min) and stimulated subsequently with LPS (1 µg/ml) for 20 h. Levels as well as gene expressions of various cytokines were compared with that of LPS alone incubated cells. Alongside, in vivo oral anti-inflammatory efficacy against LPS induced endotoxemia study was effectuated, wherein rats were administered with CETC 48, 24, 12 and 1 h prior to the injection of LPS and the survival of rats were monitored upto 10 days. Cytokine levels were quantified by ELISA. Nitrite levels were measured using Griess reagent. Expression of pro-inflammatory proteins was inspected in rat tissues by histochemical and immuno -histochemical examinations. RESULTS: CETC was able to down-regulate the up-regulation of pro-inflammatory biomarkers in THP-1 macrophages though blockade of NF-κB nuclear translocation and could improve the survival rate during endotoxemic episodes with a marked suppression of the tissue expression of pro-inflammatory proteins. CONCLUSION: These findings concomitantly reveal the anti-inflammatory mechanism of CETC and support us to move forward for the development of drugs against disorders resulting from deregulated immune reactions.
Assuntos
Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Sepse/tratamento farmacológico , Tinospora/química , Animais , Citocinas/metabolismo , Avaliação Pré-Clínica de Medicamentos , Endotoxemia/tratamento farmacológico , Humanos , Índia , Inflamação/tratamento farmacológico , Lipopolissacarídeos , Masculino , NF-kappa B/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Células THP-1RESUMO
BACKGROUND: The ongoing treatment modalities for breast cancer (BC) primarily rely on the expression status of ER, PR and HER-2 receptors in BC tissues. Our strategy of chemosensitization provides new insights to counter chemoresistance, a major obstacle that limits the benefits of chemotherapy of mammary cancers. METHODS: By utilizing a murine breast cancer model employing NSG mice bearing orthotopic triple-negative breast cancer (TNBC) xenografts, we have evaluated the ability of phytochemical curcumin in chemosensitizing BC to 5-Fluorouracil (5-FU) chemotherapy and the differential modulations of cellular events in response to this strategy, independent of their receptor status. RESULTS: A significant synergistic antitumor potential was observed in the murine model with a sub-optimal dose treatment of 5-FU plus curcumin, as evaluated by a reduction in the tumor-related parameters. We authenticated the pivotal role of thymidylate synthase (TS) in regulating the 5-FU-curcumin synergism using the TNBC pre-clinical model. Our study also confirmed the pharmacological safety of this chemotherapeutic plus phytoactive combination using acute and chronic toxicity studies in Swiss albino mice. Subsequently, the molecular docking analysis of curcumin binding to TS demonstrated the affinity of curcumin towards the cofactor-binding site of TS, rather than the substrate-binding site, where 5-FU binds. Our concomitant in vivo and in silico evidence substantiates the superior therapeutic index of this combination. CONCLUSION: This is the first-ever pre-clinical study portraying TS as the critical target of combinatorial therapy for mammary carcinomas and therefore we recommend its clinical validation, especially in TNBC patients, who currently have limited therapeutic options.
RESUMO
OBJECTIVE: Clinical trials have demonstrated the efficacy of indigo naturalis, a traditional Chinese medicine ingredient, against psoriasis, a skin disease characterized by keratinocyte hyperproliferation and inflammation. The present study investigates the efficacy of tryptanthrin, a bioactive compound in indigo naturalis, against non-melanoma skin cancer (NMSC) and the signalling events involved. METHODS: Efficacy of tryptanthrin against NMSC was assessed using DMBA/PMA-induced skin carcinogenesis model in Swiss albino mice. Immunostaining for PCNA and ki-67 was used to mark proliferating cells in tissues. Haematoxylin and eosin staining and toluidine staining were employed to assess inflammation, and TUNEL assay was used to detect apoptosis in tissues. The signalling events were evaluated using Western blot, imunohistochemistry and immunofluorescence staining. MTT assay and clonogenic assay were performed to assess the viability and proliferation of cancer cells, in vitro. RESULTS: In mice, topical application of tryptanthrin suppressed skin carcinogenesis. It attenuated inflammation, impeded the proliferation of hair follicle (HF) cells and suppressed the activation of ß-catenin, a major driver of HF cell proliferation. Additionally tryptanthrin suppressed the activation of ERK1/2 and p38, both of which promote ß-catenin activation and lowered the expression of c-Myc and cyclin-D1. Tryptanthrin suppressed the proliferation of the human NMSC cell line, A431 and abrogated EGF-induced activation of ß-catenin and subsequent cytoskeletal rearrangement. CONCLUSION: The study demonstrates with molecular evidence that tryptanthrin is an effective suppressor of NMSC.
Assuntos
Quinazolinas/farmacologia , Neoplasias Cutâneas/prevenção & controle , Animais , Ensaios de Seleção de Medicamentos Antitumorais , Antígeno Ki-67/metabolismo , Camundongos , Proteínas de Neoplasias/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Neoplasias Cutâneas/induzido quimicamente , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Acetato de Tetradecanoilforbol/toxicidade , Ácido gama-Aminobutírico/análogos & derivados , Ácido gama-Aminobutírico/toxicidadeRESUMO
An amendment to this paper has been published and can be accessed via a link at the top of the paper.