RESUMEN
Spisulosine (1-deoxysphinganine) is a sphingoid amino alcohol isolated from the sea clams that showed potent antiproliferative activity against a broad spectrum of solid tumors but failed in clinical trials due to neurotoxicity. However, its structural similarity to other bioactive sphingoids, interesting mode of action, and appreciable potency against cancer cells make it a suitable lead for future anticancer drug development. The present study was conducted to elucidate mechanisms of the antiproliferative/cytotoxic effects of newly synthesized spisulosine analog homospisulosine (KP7). The evaluation was performed on cervical carcinoma cells, representing an in vitro model of one of the most common cancer types and a significant worldwide cause of women's cancer mortality. Treatment with homospisulosine (2.0 µM) for 24, 48, and 72 h significantly inhibited the growth of HeLa cells in vitro and induced apoptosis detectable by DNA fragmentation, externalization of phosphatidylserine, dissipation of mitochondrial membrane potential, activation of caspase-3 and cleavage of PARP. In addition, treating HeLa cells with spisulosine increased p27 and Bcl-2 on protein levels and phosphorylation of Bcl-2 on Ser70 residue. These results support the potential for spisulosine analogs represented here by homospisulosine for future therapeutic development.
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Antineoplásicos , Carcinoma , Femenino , Humanos , Células HeLa , Regulación hacia Arriba , Fosforilación , Apoptosis , Antineoplásicos/farmacologíaRESUMEN
Tick-borne encephalitis virus (TBEV) is transmitted mainly by tick bites, but humans can acquire infection through consuming unpasteurized milk from infected animals. Interhuman transmission of TBEV by breast milk has not been confirmed or ruled out. We report a case of probable transmission of TBEV from an unvaccinated mother to an infant through breast-feeding.
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Virus de la Encefalitis Transmitidos por Garrapatas , Encefalitis Transmitida por Garrapatas , Mordeduras de Garrapatas , Animales , Femenino , Humanos , Leche Humana , MadresRESUMEN
Non-pharmaceutical interventions (NPIs) were the only viable choice to mitigate or suppress transmission of COVID-19 in the absence of efficient and safe vaccines. In this study, we examined the association between the stringency of containment measures and cumulative incidence of the COVID-19 cases in the first wave of the pandemic across 28 European countries. Our results support the effectiveness of containment measures in the mitigation or suppression of COVID-19 epidemics. Early adoption of stringent containment measures prior to detection of the first confirmed case, together with ramping up containment stringency during the early days of epidemics, was associated with a lower disease occurrence. The delayed adoption of stringent containment measures did not fully compensate for the lack of early response. Containment measures continue to play a significant role in the control of COVID-19 in the post-vaccination period, when limited vaccination coverage, the emergence of vaccine resistance, and/or increased mobility enabled further disease transmission (Tab. 4, Fig. 22, Ref. 50). Keywords: non-pharmaceutical interventions, containment, COVID-19, mobility, social distancing, Containment and Health Index, epidemiology, public health measures, SARS-CoV-2.
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COVID-19 , Control de Enfermedades Transmisibles , Cuarentena , COVID-19/epidemiología , COVID-19/prevención & control , Control de Enfermedades Transmisibles/métodos , Europa (Continente) , Humanos , Incidencia , Pandemias/prevención & control , SARS-CoV-2RESUMEN
Carcinogenicity of hexavalent chromium [Cr (VI)] has been supported by a number of epidemiological and animal studies; however, its carcinogenic mode of action is still incompletely understood. To identify mechanisms involved in cancer development, we analyzed gene expression data from duodena of mice exposed to Cr(VI) in drinking water. This analysis included (i) identification of upstream regulatory molecules that are likely responsible for the observed gene expression changes, (ii) identification of annotated gene expression data from public repositories that correlate with gene expression changes in duodena of Cr(VI)-exposed mice, and (iii) identification of hallmark and oncogenic signature gene sets relevant to these data. We identified the inactivated CFTR gene among the top scoring upstream regulators, and found positive correlations between the expression data from duodena of Cr(VI)-exposed mice and other datasets in public repositories associated with the inactivation of the CFTR gene. In addition, we found enrichment of signatures for oncogenic signaling, sustained cell proliferation, impaired apoptosis and tissue remodeling. Results of our computational study support the tumor-suppressor role of the CFTR gene. Furthermore, our results support human relevance of the Cr(VI)-mediated carcinogenesis observed in the small intestines of exposed mice and suggest possible groups that may be more vulnerable to the adverse outcomes associated with the inactivation of CFTR by hexavalent chromium or other agents. Lastly, our findings predict, for the first time, the role of CFTR inactivation in chemical carcinogenesis and expand the range of plausible mechanisms that may be operative in Cr(VI)-mediated carcinogenesis of intestinal and possibly other tissues.
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Transformación Celular Neoplásica/inducido químicamente , Cromo/toxicidad , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Neoplasias Duodenales/inducido químicamente , Duodeno/efectos de los fármacos , Silenciador del Gen/efectos de los fármacos , Proteínas Supresoras de Tumor/genética , Contaminantes Químicos del Agua/toxicidad , Administración Oral , Animales , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/metabolismo , Transformación Celular Neoplásica/patología , Cromo/administración & dosificación , Regulador de Conductancia de Transmembrana de Fibrosis Quística/metabolismo , Bases de Datos Genéticas , Agua Potable , Neoplasias Duodenales/genética , Neoplasias Duodenales/metabolismo , Neoplasias Duodenales/patología , Duodeno/metabolismo , Duodeno/patología , Perfilación de la Expresión Génica , Ratones , Medición de Riesgo , Biología de Sistemas , Transcriptoma , Proteínas Supresoras de Tumor/metabolismo , Contaminantes Químicos del Agua/administración & dosificaciónRESUMEN
Oncogenic gene fusions represent attractive targets for therapy of cancer. However, the frequency of actionable genomic rearrangements in colorectal cancer (CRC) is very low, and universal screening for these alterations seems to be impractical and costly. To address this problem, several large scale studies retrospectivelly showed that CRC with gene fusions are highly enriched in groups of tumors defined by MLH1 DNA mismatch repair protein deficiency (MLH1d), and hypermethylation of MLH1 promoter (MLH1ph), and/or the presence of microsatellite instability, and BRAF/KRAS wild-type status (BRAFwt/KRASwt). In this study, we used targeted next generation sequencing (NGS) to explore the occurence of potentially therapeutically targetable gene fusions in an unselected series of BRAFwt/KRASwt CRC cases that displayed MLH1d/MLH1ph. From the initially identified group of 173 MLH1d CRC cases, 141 cases (81.5%) displayed MLH1ph. BRAFwt/RASwt genotype was confirmed in 23 of 141 (~16%) of MLH1d/MLH1ph cases. Targeted NGS of these 23 cases identified oncogenic gene fusions in nine patients (39.1%; CI95: 20.5%-61.2%). Detected fusions involved NTRK (four cases), ALK (two cases), and BRAF genes (three cases). As a secondary outcome of NGS testing, we identified PIK3K-AKT-mTOR pathway alterations in two CRC cases, which displayed PIK3CA mutation. Altogether, 11 of 23 (~48%) MLH1d/MLH1ph/BRAFwt/RASwt tumors showed genetic alterations that could induce resistance to anti-EGFR therapy. Our study confirms that targeted NGS of MLH1d/MLH1ph and BRAFwt/RASwt CRCs could be a cost-effective strategy in detecting patients with potentially druggable oncogenic kinase fusions.
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Adenocarcinoma/genética , Neoplasias Colorrectales/genética , Pruebas Genéticas/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Homólogo 1 de la Proteína MutL/deficiencia , Proteínas de Fusión Oncogénica/genética , Adenocarcinoma/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Quinasa de Linfoma Anaplásico/genética , Neoplasias Colorrectales/diagnóstico , Metilación de ADN , Femenino , Pruebas Genéticas/normas , Secuenciación de Nucleótidos de Alto Rendimiento/normas , Humanos , Masculino , Persona de Mediana Edad , Homólogo 1 de la Proteína MutL/genética , Regiones Promotoras Genéticas , Proteínas Proto-Oncogénicas B-raf/genética , Proteínas Proto-Oncogénicas p21(ras)/genética , Proteínas Tirosina Quinasas Receptoras/genéticaRESUMEN
High throughput screening (HTS) and functional genomics (toxicogenomics) have opened new avenues in toxicity testing. Their advantages include the potential for developing short-term in vivo bioassays and in vitro assays in order to keep pace with the growing backlog of chemicals that need to be evaluated for potential human health risk. In addition, these approaches have the potential to address some of the difficulties that arise with interpreting traditional rodent bioassays, such as the relevance of apical outcomes induced by chemical exposure in animals to humans. The wealth of information associated with the HTS and toxicogenomic data can inform human health risk assessment primarily through (i) insight into potential mechanism of action, (ii) prediction of adverse outcomes of chemical exposures, and (iii) dose-response assessment for derivation of toxicity values. In this article we outline current and expected future progress in these three directions and argue for increased role of HTS and toxicogenomic data in chemical risk assessment. We conclude that these approaches can help fulfill the NRC vision for toxicity testing in the 21st century and we discuss specific examples of chemicals whose health assessments can potentially benefit from available HTS or toxicogenomic data.
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Ensayos Analíticos de Alto Rendimiento , Medición de Riesgo/métodos , Toxicogenética/métodos , Animales , Carcinógenos/toxicidad , Humanos , TranscriptomaRESUMEN
We present macroscopic and microscopic findings in a case of chronic dissecting aneurysm of ascending aorta and aortic arch associated with isolated tears of aortic wall without its rupture in a 71-year-old female presenting with minimal clinical symptomatology. Aneurysmal dilation of the aorta was caused not only by the bridging of the vascular wall based on the presence of an organizing intramural thrombus in the false lumen between the separated layers, but also by a wide flat defect in the aortic intima and media with the preservation of the aortic wall integrity due to fibrotical alteration of tunica adventicia. Histologic examination of the thoracic aorta detected cystic medial degeneration with mild atherosclerosis.
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Disección Aórtica , Trombosis , Anciano , Disección Aórtica/diagnóstico , Aorta/anomalías , Femenino , Humanos , Trombosis/diagnósticoRESUMEN
Pulmonary small cell carcinoma (SCLC) is usually diagnosed in small biopsy or cytological specimens based on cytomorphology; however in ambiguous cases diagnosis requires additional support by immunohistochemistry. While TP53 and RB1 alterations with secondary overexpression of p16 are mainstay events in SCLC pathogenesis, diagnostic value of p16-positivity in the diagnosis of SCLC has not yet been fully investigated. We examined the expression of p16, CD56, synaptophysin (SYP), chromogranin A and thyroid transcription factor-1 (TTF1) in a series of pulmonary and extrapulmonary small cell carcinomas, pulmonary carcinoids and non-small cell lung carcinomas, and compared diagnostic performance of these markers in the diagnosis of SCLC. P16 was positive in 95 of 101 SCLCs, and displayed highest diagnostic sensitivity of ~94%. Composite biomarkers CD56+p16+TTF1 and CD56+p16+SYP were both able to detect correctly all SCLC cases. Importantly, three (~3%) SCLC cases completely negative for all conventional markers displayed diffuse positivity for p16. CD56 and p16 demonstrated highest concordance between paired small biopsy and cytology specimens. 50% of squamous cell carcinomas, ~41% of adenocarcinoma/NSCLC-favour adenocarcinoma cases, and ~93% of extrapulmonary small cell carcinomas also showed p16-positivity. Combination of CD56, p16 and TTF1 produced diagnostic classifier that outperformed best single marker CD56 in differential diagnosis between SCLC and NSCLC. In conclusion, in the appropriate morphological context p16 represents a useful supplementary marker for diagnosis of SCLC, even in cases where only cytological material is available.
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Biomarcadores de Tumor/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Carcinoma de Células Pequeñas/patología , Genes p16/fisiología , Neoplasias Pulmonares/patología , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Tumor Carcinoide/diagnóstico , Tumor Carcinoide/patología , Carcinoma de Células Pequeñas/diagnóstico , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patología , Diagnóstico Diferencial , Femenino , Humanos , Inmunohistoquímica/métodos , Pulmón/patología , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: Epithelial-to-mesenchymal transition (EMT) has been associated with the acquisition of metastatic potential and the resistance of cancer cells to therapeutic treatments. MCF-7 breast cancer cells engineered to constitutively express the zinc-finger transcriptional repressor gene Snail (MCF-7-Snail cells) have been previously shown to display morphological and molecular changes characteristic of EMT. We report here the results of a comprehensive systems level molecular analysis of changes in global patterns of gene expression and levels of glutathione and reactive oxygen species (ROS) in MCF-7-Snail cells and the consequence of these changes on the sensitivity of cells to radiation treatment and therapeutic drugs. METHODS: Snail-induced changes in global patterns of gene expression were identified by microarray profiling using the Affymetrix platform (U133 Plus 2.0). The resulting data were processed and analyzed by a variety of system level analytical methods. Levels of ROS and glutathione (GSH) were determined by fluorescent and luminescence assays, and nuclear levels of NF-κB protein were determined by an ELISA based method. The sensitivity of cells to ionizing radiation and anticancer drugs was determined using a resazurin-based cell cytotoxicity assay. RESULTS: Constitutive ectopic expression of Snail in epithelial-like, luminal A-type MCF-7 cells induced significant changes in the expression of >7600 genes including gene and miRNA regulators of EMT. Mesenchymal-like MCF-7-Snail cells acquired molecular profiles characteristic of triple-negative, claudin-low breast cancer cells, and displayed increased sensitivity to radiation treatment, and increased, decreased or no change in sensitivity to a variety of anticancer drugs. Elevated ROS levels in MCF-7-Snail cells were unexpectedly not positively correlated with NF-κB activity. CONCLUSIONS: Ectopic expression of Snail in MCF-7 cells resulted in morphological and molecular changes previously associated with EMT. The results underscore the complexity and cell-type dependent nature of the EMT process and indicate that EMT is not necessarily predictive of decreased resistance to radiation and drug-based therapies.
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Neoplasias de la Mama/genética , Transición Epitelial-Mesenquimal/genética , Proteínas de Neoplasias/biosíntesis , Factores de Transcripción de la Familia Snail/biosíntesis , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Neoplasias de la Mama/radioterapia , Resistencia a Antineoplásicos/genética , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Humanos , Células MCF-7 , FN-kappa B/biosíntesis , FN-kappa B/genética , Proteínas de Neoplasias/genética , Tolerancia a Radiación/genética , Especies Reactivas de Oxígeno/metabolismo , Factores de Transcripción de la Familia Snail/genéticaRESUMEN
A growing body of evidence suggests that the developmental process of epithelial-to-mesenchymal transition (EMT) is co-opted by cancer cells to metastasize to distant sites. This transition is associated with morphologic elongation and loss of cell-cell adhesions, though little is known about how it alters cell biophysical properties critical for migration. Here, we use multiple-particle tracking (MPT) microrheology and traction force cytometry to probe how genetic induction of EMT in epithelial MCF7 breast cancer cells changes their intracellular stiffness and extracellular force exertion, respectively, relative to an empty vector control. This analysis demonstrated that EMT alone was sufficient to produce dramatic cytoskeletal softening coupled with increases in cell-exerted traction forces. Microarray analysis revealed that these changes corresponded with down-regulation of genes associated with actin cross-linking and up-regulation of genes associated with actomyosin contraction. Finally, we show that this loss of structural integrity to expedite migration could inhibit mesenchymal cell proliferation in a secondary tumor as it accumulates solid stress. This work demonstrates that not only does EMT enable escape from the primary tumor through loss of cell adhesions but it also induces a concerted series of biophysical changes enabling enhanced migration of cancer cells after detachment from the primary tumor.
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Citoesqueleto/genética , Transición Epitelial-Mesenquimal/genética , Transición Epitelial-Mesenquimal/fisiología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Actinas/metabolismo , Fenómenos Biofísicos , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Núcleo Celular/metabolismo , Núcleo Celular/patología , Citoesqueleto/patología , Femenino , Expresión Génica , Humanos , Células MCF-7 , Análisis de Secuencia por Matrices de Oligonucleótidos , Fenotipo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Factores de Transcripción de la Familia Snail , Transformación GenéticaRESUMEN
An increased rate of second nonmelanoma skin cancers is found in immunocompromised patients. Epidemiological and molecular data implicate ultraviolet radiation as the major risk factor. In addition, there is increasing evidence supporting the role of human papillomavirus (HPV) in the pathogenesis of premalignant and malignant skin lesions in both immunocompetent and immunocompromised patients. In a retrospective cross-sectional study, the authors examined the expression of p16 by immunohistochemistry and the presence of mucosal (α-genus) and cutaneous/epidermodysplasia verruciformis (ß-genus) HPV DNA by polymerase chain reaction in 29 biopsy specimens of extragenital/extraungual Bowen disease (BD) from 24 Eastern European white immunocompromised patients. Furthermore, the author evaluated the association between the expression of p16 protein and the presence of HPV DNA. Among 25 specimens from 21 patients evaluable by polymerase chain reaction, HPV DNA was detected in 10 (40%) BD lesions from 9 patients. Beta-HPV predominated over alpha-HPV types. Among 29 immunohistochemically evaluable BD specimens, 22 lesions (â¼76%) from 20 patients were scored as p16 positive. HPV DNA-positive and HPV DNA-negative lesions displayed the same proportion of p16 positivity (80%) and no correlation was found between the HPV DNA presence and the p16 expression status. Our pilot study demonstrated that ß-HPV infections predominate in BD cases diagnosed among immunocompromised patients, although high- and low-risk mucosal (alpha) HPV genotypes may be detected in a minority of cases. In contrast to anogenital HPV-associated lesions, positive p16 expression is not a reliable marker of high-risk α-HPV infection in BD cases, as it can be also detected in ß-HPV infected and HPV-negative cases.
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Biomarcadores de Tumor/análisis , Enfermedad de Bowen/virología , Inhibidor p16 de la Quinasa Dependiente de Ciclina/análisis , Huésped Inmunocomprometido , Papillomaviridae/patogenicidad , Infecciones por Papillomavirus/virología , Lesiones Precancerosas/virología , Neoplasias Cutáneas/virología , Anciano , Anciano de 80 o más Años , Biopsia , Enfermedad de Bowen/química , Enfermedad de Bowen/inmunología , Transformación Celular Viral , Estudios Transversales , República Checa , ADN Viral/genética , Femenino , Pruebas de ADN del Papillomavirus Humano , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Papillomaviridae/genética , Papillomaviridae/inmunología , Infecciones por Papillomavirus/inmunología , Proyectos Piloto , Lesiones Precancerosas/química , Lesiones Precancerosas/inmunología , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Factores de Riesgo , Neoplasias Cutáneas/química , Neoplasias Cutáneas/inmunologíaRESUMEN
Human papillomavirus (HPV)-associated oropharyngeal squamous cell carcinoma (HPV-OSCC) represents a specific clinical and pathological entity among head and neck cancers with more favorable prognosis than corresponding HPV-negative oropharyngeal squamous cell carcinoma (SCC). HPV-OSCC most commonly displays non-keratinzing SCC histology, although number of cases presented variable morphology. Detection of transcriptionally active HPV in oropharyngeal SCCs is of critical prognostic importance, which even supersedes the importance of their formal pathological grading. This article summarizes the current state of knowledge of HPV-positive SCC of oropharynx and of other anatomical subsites of the head and neck and discusses the role of HPV detection in the diagnostics of metastatic SCC lesions of unknown origin.
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Carcinoma de Células Escamosas/patología , Neoplasias de Cabeza y Cuello/patología , Papillomaviridae , Infecciones por Papillomavirus/patología , Carcinoma de Células Escamosas/virología , Neoplasias de Cabeza y Cuello/virología , Humanos , Inmunohistoquímica , Hibridación in Situ , Infecciones por Papillomavirus/virología , PronósticoRESUMEN
Prostate cancer (PCa) is the second leading cause of cancer-related mortality in men. The prevalent diagnosis method is based on the serum prostate-specific antigen (PSA) screening test, which suffers from low specificity, overdiagnosis, and overtreatment. In this work, untargeted metabolomic profiling of age-matched serum samples from prostate cancer patients and healthy individuals was performed using ultraperformance liquid chromatography coupled to high-resolution tandem mass spectrometry (UPLC-MS/MS) and machine learning methods. A metabolite-based in vitro diagnostic multivariate index assay (IVDMIA) was developed to predict the presence of PCa in serum samples with high classification sensitivity, specificity, and accuracy. A panel of 40 metabolic spectral features was found to be differential with 92.1% sensitivity, 94.3% specificity, and 93.0% accuracy. The performance of the IVDMIA was higher than the prevalent PSA test. Within the discriminant panel, 31 metabolites were identified by MS and MS/MS, with 10 further confirmed chromatographically by standards. Numerous discriminant metabolites were mapped in the steroid hormone biosynthesis pathway. The identification of fatty acids, amino acids, lysophospholipids, and bile acids provided further insights into the metabolic alterations associated with the disease. With additional work, the results presented here show great potential toward implementation in clinical settings.
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Biomarcadores de Tumor/sangre , Neoplasias de la Próstata/diagnóstico , Anciano , Biomarcadores de Tumor/aislamiento & purificación , Estudios de Casos y Controles , Cromatografía Líquida de Alta Presión , Estudios de Factibilidad , Humanos , Masculino , Metabolómica , Persona de Mediana Edad , Neoplasias de la Próstata/sangre , Espectrometría de Masas en TándemRESUMEN
OBJECTIVE: We recently determined that the ectopic over-expression of miR-429 and other members of the miR-200 family of microRNAs in ovarian cancer (OC) mesenchymal-like cell lines induces mesenchymal-to-epithelial transition (MET) with a concomitant increase in sensitivity to platinum drugs. We sought to determine if metastasizing OC cells isolated from an OC patient could also be induced by miR-429 to undergo MET and become sensitized to established first-line platinum-based therapies. METHODS: We established and characterized a new primary cell line (OCI-984) from free-floating OC cells isolated from the ascites fluid of an advanced stage OC patient. miR-429 was ectopically over-expressed in these cells. RESULTS: The over-expression of miR-429 in OCI-984 cells induced morphological, functional and molecular changes consistent with MET and a concomitant significant increase in the sensitivity of the converted cells to cisplatin. CONCLUSIONS: Our findings indicate that the miR-200 family of microRNAs, and miR-429 in particular, play a critical role in the functioning of OC metastasizing cells and that targeted delivery of miR-429, and perhaps other miR-200 family members, in combination with platinum-based chemotherapies may be an effective strategy in reducing OC metastasis and tumor recurrence.
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Antineoplásicos/farmacología , Línea Celular Tumoral , Cisplatino/farmacología , MicroARNs/biosíntesis , Neoplasias Ováricas/genética , Neoplasias Ováricas/patología , Anciano , Animales , Ensayos de Selección de Medicamentos Antitumorales , Transición Epitelial-Mesenquimal/genética , Femenino , Humanos , Ratones , Ratones Desnudos , Neoplasias Ováricas/tratamiento farmacológico , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Camalexin, the phytoalexin produced in the model plant Arabidopsis thaliana, possesses antiproliferative and cancer chemopreventive effects. We have demonstrated that the cytostatic/cytotoxic effects of camalexin on several prostate cancer (PCa) cells are due to oxidative stress. Lysosomes are vulnerable organelles to Reactive Oxygen Species (ROS)-induced injuries, with the potential to initiate and or facilitate apoptosis subsequent to release of proteases such as cathepsin D (CD) into the cytosol. We therefore hypothesized that camalexin reduces cell viability in PCa cells via alterations in expression and activity of CD. Cell viability was evaluated by MTS cell proliferation assay in LNCaP and ARCaP Epithelial (E) cells, and their respective aggressive sublines C4-2 and ARCaP Mesenchymal (M) cells, whereby the more aggressive PCa cells (C4-2 and ARCaPM) displayed greater sensitivity to camalexin treatments than the lesser aggressive cells (LNCaP and ARCaPE). Immunocytochemical analysis revealed CD relocalization from the lysosome to the cytosol subsequent to camalexin treatments, which was associated with increased protein expression of mature CD; p53, a transcriptional activator of CD; BAX, a downstream effector of CD, and cleaved PARP, a hallmark for apoptosis. Therefore, camalexin reduces cell viability via CD and may present as a novel therapeutic agent for treatment of metastatic prostate cancer cells.
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Antineoplásicos Fitogénicos/farmacología , Arabidopsis/química , Catepsina D/genética , Regulación de la Expresión Génica/efectos de los fármacos , Indoles/farmacología , Tiazoles/farmacología , Antineoplásicos Fitogénicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Catepsina D/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Humanos , Indoles/aislamiento & purificación , Lisosomas/efectos de los fármacos , Lisosomas/metabolismo , Masculino , Especificidad de Órganos , Estrés Oxidativo , Extractos Vegetales/química , Poli(ADP-Ribosa) Polimerasas/genética , Poli(ADP-Ribosa) Polimerasas/metabolismo , Próstata/efectos de los fármacos , Próstata/enzimología , Próstata/patología , Transducción de Señal , Tiazoles/aislamiento & purificación , 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
Absolute (ALW) and relative (RLW) liver weight changes are sensitive endpoints in repeat-dose rodent toxicity studies, and their changes are often used for quantitative assessment of health effects induced by hepatotoxic chemicals using the benchmark dose-response modeling (BMD) approach. To find biologically relevant liver weight changes to chemical exposures, we evaluated all data available for liver weight changes and associated liver histopathologic findings from the Toxicity Reference Database (ToxRefDB). Our analysis of 389 subchronic mouse and rat studies for 273 chemicals found significant differences in treatment-related ALW and RLW changes between dose groups with and without liver histopathologic changes. In addition, we demonstrate that chemical treatment-induced ALW and RLW changes can predict the presence of histopathologic findings and inform the selection of biologically relevant liver weight changes for BMD modeling and derivation of toxicity values.
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Enfermedad Hepática Inducida por Sustancias y Drogas , Bases de Datos Factuales , Hígado , Animales , Hígado/efectos de los fármacos , Hígado/patología , Ratones , Ratas , Tamaño de los Órganos/efectos de los fármacos , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Relación Dosis-Respuesta a Droga , MasculinoRESUMEN
Straightforward access to enantiomerically pure 3,4-diamino-3,4-dideoxyphytosphingosines, as novel analogues of natural d-ribo-phytosphingosine was accomplished, starting from two available chirons: dimethyl l-tartrate and d-isoascorbic acid. A sequential Overman rearrangement followed by late-stage introduction of the alkyl side chain moiety via olefin cross-metathesis is the cornerstone of this approach. The preliminary evaluation study of the synthesised sphingomimetics, based on their ability to inhibit a proliferation of human cancer cells, showed promising cytotoxicity against Jurkat and HeLa cells for (2R,3R,4S)-2,3,4-triaminooctadecan-1-ol trihydrochloride.
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Proliferación Celular , Esfingosina , Esfingosina/análogos & derivados , Esfingosina/química , Esfingosina/farmacología , Esfingosina/síntesis química , Humanos , Células HeLa , Proliferación Celular/efectos de los fármacos , Células Jurkat , Antineoplásicos/farmacología , Antineoplásicos/química , Antineoplásicos/síntesis química , EstereoisomerismoRESUMEN
Preliminary studies have suggested that the reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine (NAC) may be effective in inhibiting the growth of pancreatic cancer cells. In-depth cellular and molecular analyses were carried out to determine NAC's mode of action in inhibiting the growth of a well-characterized pancreatic cancer cell line (AsPC-1). Standardized assays were used to monitor cellular growth, apoptosis, levels of ROS, cellular senescence, migration, and invasiveness. Cell stiffness was measured using atomic force microscopy. Gene expression was monitored by quantitative PCR. NAC significantly inhibits the growth and metastatic potential of AsPC-1 cells by inducing cell-cycle arrest in G1 and subsequent cellular senescence and decreased invasiveness. These anticancer properties are associated with an unexpected increase in the intracellular concentrations of ROS. NAC does not decrease the susceptibility of AsPC-1 cells to the anticancer drugs gemcitabine, mitomycin C, and doxorubicin. NAC-induced changes in gene expression are consistent with the onset of mesenchymal-to-epithelial transition. In conclusion, our findings indicate that NAC induces an integrated series of responses in AsPC-1 cells that make it a highly promising candidate for development as a pancreatic cancer therapeutic.
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Acetilcisteína/farmacología , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/patología , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Senescencia Celular/efectos de los fármacos , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacología , Relación Dosis-Respuesta a Droga , Doxorrubicina/farmacología , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Genes myc , Humanos , Mitomicina/farmacología , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , GemcitabinaRESUMEN
An effective synthesis of analogs of the indole phytoalexin cyclobrassinin with NR1R2 group instead of SCH3 was developed starting from indole-3-carboxaldehyde. The target compounds were prepared by spirocyclization of 1-Boc-thioureas with the formation of isolable spiroindoline intermediates, followed by the trifluoroacetic acid-induced cascade reaction consisting of methanol elimination, deprotection and rearrangement of the iminium ion. The structures of novel products were elucided by the (1)H and (13)C NMR spectroscopy, including HMBC, HSQC, COSY, NOESY and DEPT measurements. Several newly synthesized compounds demonstrated significant antiproliferative/cytotoxic activity against human leukemia and solid tumor cell lines, as well as remarkable selectivity of these effects against cancer cells relative to the non-malignant HUVEC cells.
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Antineoplásicos/síntesis química , Indoles/química , Sesquiterpenos/química , Tiocarbamatos/química , Antineoplásicos/química , Antineoplásicos/toxicidad , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Células HeLa , Células Endoteliales de la Vena Umbilical Humana , Humanos , Indoles/síntesis química , Indoles/toxicidad , Células Jurkat , Células MCF-7 , Espectroscopía de Resonancia Magnética , Estereoisomerismo , Tiocarbamatos/síntesis química , Tiocarbamatos/toxicidad , FitoalexinasRESUMEN
Programmed cell death plays a crucial role in maintaining the homeostasis and integrity of multicellular organisms, and its dysregulation contributes to the pathogenesis of many diseases. Programmed cell death is regulated by a range of macromolecules and low-molecular messengers, including ceramides. Endogenous ceramides have different functions, that are influenced by their localization and the presence of their target molecules. This article provides an overview of the current understanding of ceramides and their impact on various types of programmed cell death, including apoptosis, anoikis, macroautophagy and mitophagy, and necroptosis. Moreover, it highlights the emergence of dihydroceramides as a new class of bioactive sphingolipids and their downstream targets as well as their future roles in cancer cell growth, drug resistance and tumor metastasis.