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1.
BMC Cancer ; 19(1): 994, 2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31646972

RESUMO

BACKGROUND: Long non coding RNAs (lncRNAs) are RNA molecules longer than 200 nucleotides that are not translated into proteins, but regulate the transcription of genes involved in different cellular processes, including cancer. Epidemiological analyses have demonstrated that parous women have a decreased risk of developing breast cancer in postmenopausal years if they went through a full term pregnancy in their early twenties. We here provide evidence of the role of BC200 in breast cancer and, potentially, in pregnancy's preventive effect in reducing the lifetime risk of developing breast cancer. METHODS: Transcriptome analysis of normal breast of parous and nulliparous postmenopausal women revealed that several lncRNAs are differentially expressed in the parous breast. RNA sequencing of healthy postmenopausal breast tissue biopsies from eight parous and eight nulliparous women showed that there are 42 novel lncRNAs differentially expressed between these two groups. Screening of several of these 42 lncRNAs by RT-qPCR in different breast cancer cell lines, provided evidence that one in particular, lncEPCAM (more commonly known as BC200), was a strong candidate involved in cancer progression. Proliferation, migration, invasion and xerograph studies confirmed this hypothesis. RESULTS: The poorly studied oncogenic BC200 was selected to be tested in vitro and in vivo to determine its relevance in breast cancer and also to provide us with an understanding of its role in the increased susceptibility of the nulliparous women to cancer. Our results show that BC200 is upregulated in nulliparous women, and breast cancer cells and tissue. The role of BC200 is not completely understood in any of the breast cancer subtypes. We here provide evidence that BC200 has a role in luminal breast cancer as well as in the triple negative breast cancer subtype. CONCLUSION: When overexpressed in luminal and triple negative breast cancer cell lines, BC200 shows increased proliferation, migration, and invasion in vitro. In vivo, overexpression of BC200 increased tumor size. Although treatment for cancer using lncRNAs as targets is in its infancy, the advancement in knowledge and technology to study their relevance in disease could lead to the development of novel treatment and preventive strategies for breast cancer.


Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/patologia , RNA Longo não Codificante/genética , Animais , Apoptose , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Progressão da Doença , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Camundongos SCID , Transplante de Neoplasias , Paridade , Pós-Menopausa , Gravidez , RNA Longo não Codificante/metabolismo , Receptores Estrogênicos/metabolismo , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologia , Carga Tumoral , Regulação para Cima
2.
Int J Mol Sci ; 20(18)2019 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-31546809

RESUMO

This paper aims to apply a proposed, based on calorimetric measurements, a reliable numerical model for magnetic fluid hyperthermia (MFH) treatment planning of breast cancer. Furthermore, we perform a comparative analysis of magnetic nanoparticles (MNPs) and tumour tissue interactions by means of the magnetic-field-dependent Néel and Brownian relaxation times. The analysis was based on an anatomically correct breast model (developed in-house) and a modified linear response theory, which was applied to investigate the heat dissipation from the magnetic nanoparticles dispersed in the breast tumour. The calculations of the single-domain magnetic power losses were conducted for a case where the magnetic field value and the applied frequency were known, but also for the different concentrations of the MNPs in the tumour. Two scenarios were considered: The MNPs mobilised and immobilised in the tumour. In parallel, the eddy currents effect, together with the related temperature distributions, were considered in order to analyse safety issues. By changing the MNP concentration in the tumour, the corresponding temperature distributions were calculated. The eddy current effect, together with the related temperature distribution, were considered in order to analyse safety issues. Varying the MNP concentration in the tumour, the corresponding temperature distribution was calculated. Moreover, the cumulative equivalent minutes at 43   ℃ were analysed. In the anatomically correct breast phantoms, the tissue location can lead to "hot spots" due to the eddy current effect and subsequently to the high gradients of the temperature. That is why the analysis of safety issues related to the overheating side effect should be taken into consideration during the treatment planning of magnetic fluid hyperthermia. The phenomenon of heat dissipation from MNPs is very sophisticated and depends on their concentration, the distribution and the relaxation mechanism in the tumour, together with magnetic field strength and frequency. Furthermore, we inferred that the phenomenon of heat dissipation from MNPs equally depends on MNP-tissue interactions, and it can lead to 30% differences in the power assessment. Nevertheless, the aforementioned factors should be considered in parallel using anatomical, volume-dependent models to enhance the efficiency of in vivo treatment.


Assuntos
Neoplasias da Mama , Mama , Temperatura Alta , Hipertermia Induzida , Modelos Biológicos , Imagens de Fantasmas , Mama/diagnóstico por imagem , Mama/metabolismo , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/terapia , Calorimetria , Feminino , Humanos , Nanopartículas/uso terapêutico
3.
Radiat Oncol ; 14(1): 142, 2019 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-31399108

RESUMO

BACKGROUND: Biomarkers for predicting late normal tissue toxicity to radiotherapy are necessary to personalize treatments and to optimize clinical benefit. Many radiogenomic studies have been published on this topic. Conversely, proteomics approaches are not much developed, despite their advantages. METHODS: We used the isobaric tags for relative and absolute quantitation (iTRAQ) proteomic approach to analyze differences in protein expression levels in ex-vivo irradiated (8 Gy) T lymphocytes from patients with grade ≥ 2 radiation-induced breast fibrosis (grade ≥ 2 bf+) and patients with grade < 2 bf + after curative intent radiotherapy. Patients were selected from two prospective clinical trials (COHORT and PHRC 2005) and were used as discovery and confirmation cohorts. RESULTS: Among the 1979 quantified proteins, 23 fulfilled our stringent biological criteria. Immunoblotting analysis of four of these candidate proteins (adenylate kinase 2, AK2; annexin A1; heat shock cognate 71 kDa protein; and isocitrate dehydrogenase 2) confirmed AK2 overexpression in 8 Gy-irradiated T lymphocytes from patients with grade ≥ 2 bf + compared with patients with grade < 2 bf+. As these candidate proteins are involved in oxidative stress regulation, we also evaluated radiation-induced reactive oxygen species (ROS) production in peripheral blood mononuclear cells from patients with grade ≥ 2 bf + and grade < 2 bf+. Total ROS level, and especially superoxide anion level, increased upon ex-vivo 8 Gy-irradiation in all patients. Analysis of NADPH oxidases (NOXs), a major source of superoxide ion in the cell, showed a significant increase of NOX4 mRNA and protein levels after irradiation in both patient groups. Conversely, only NOX4 mRNA level was significantly different between groups (grade ≥ 2 bf + and grade < 2 bf+). CONCLUSION: These findings identify AK2 as a potential radiosensitivity candidate biomarker. Overall, our proteomic approach highlights the important role of oxidative stress in late radiation-induced toxicity, and paves the way for additional studies on NOXs and superoxide ion metabolism.


Assuntos
Adenilato Quinase/metabolismo , Biomarcadores/metabolismo , Neoplasias da Mama/radioterapia , Mama/metabolismo , Fibrose/metabolismo , Proteoma/análise , Lesões por Radiação/metabolismo , Radioterapia/efeitos adversos , Mama/efeitos da radiação , Feminino , Fibrose/etiologia , Fibrose/patologia , Humanos , Órgãos em Risco/efeitos da radiação , Prognóstico , Estudos Prospectivos , Lesões por Radiação/etiologia , Lesões por Radiação/patologia , Tolerância a Radiação , Espécies Reativas de Oxigênio/metabolismo , Linfócitos T/metabolismo , Linfócitos T/patologia , Linfócitos T/efeitos da radiação
4.
J Steroid Biochem Mol Biol ; 194: 105447, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31415823

RESUMO

Dendrogenin A (DDA) is a newly-discovered steroidal alkaloid, which remains to date the first ever found in mammals. DDA is a cholesterol metabolites that induces cancer cell differentiation and death in vitro and in vivo, and thus behave like a tumor suppressor metabolite. Preliminary studies performed on 10 patients with estrogen receptor positive breast cancers (ER(+)BC) showed a strong decrease in DDA levels between normal matched tissue and tumors. This suggests that a deregulation on DDA metabolism is associated with breast carcinogenesis. To further investigate DDA metabolism on large cohorts of patients we have developed an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS) procedure for the quantification of DDA in liquid and in solid tissues. This method enabled the identification of DDA analogues such as its geometric isomer C17 and dendrogenin B (C26) in human samples showing that other 5,6α-epoxycholesterol conjugation products with biogenic amines exist as endogenous metabolites . We report here the first complete method of quantification of DDA in liquid and solid tissues using hydrophilic interaction liquid chromatography (HILIC). Two different methods of extraction using either a Bligh and Dyer organic extraction or protein precipitation were successfully applied to quantify DDA in solid and liquid tissues. The protein precipitation method was the fastest. The fact that this method is automatable opens up possibilities to study DDA metabolism in large cohorts of patients.


Assuntos
Colestanóis/análise , Imidazóis/análise , Mama/metabolismo , Neoplasias da Mama/metabolismo , Colestanóis/metabolismo , Cromatografia Líquida/métodos , Feminino , Humanos , Imidazóis/metabolismo
5.
J Trace Elem Med Biol ; 56: 90-99, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31442959

RESUMO

BACKGROUND: Breast cancer is the most commonly occurring neoplasm in females, comprising 16% of all female cancers worldwide. Various studies indicate some discrepancies regarding zinc (Zn) levels in various samples of breast cancer patients. OBJECTIVE: The present study evaluated by meta-analysed the published data for Zn levels analyzed in breast tissue, plasma, serum, and hair samples and its relationship with breast cancer. METHODS: The present meta-analysis included 36 studies, all of which were published in the years between 1984 to 2017 and selected by searching the databases MEDLINE, EMBASE, Cochrane Library, PubMed, Scopus, and the ISI Web of Knowledge. The articles were analyzed, and I² statistics were used to examine heterogeneity. The objective analysis was performed on data from the 36 studies, with total 1699 study subjects and 2009 controls. RESULTS: Significant statistical differences overall were observed, based on a random effects model (SMD (95 % CI), -0.78[-1.40, -0.16], P = 0.014). Data from 19 of these studies indicated significant statistical differences between cancerous patients and controls with regard to serum and plasma Zn concentration (SMD [(95 %CI): -1.61(-2.43, -0.79)]. There was a significant statistical difference between the breast tissue and hair as regards Zn status (SMD (95%CI): 2.32(1.42, 3.21)) and (SMD (95v%CI): -1.80(-3.41, -0.20), respectively. Zn concentration levels typically decreased in blood and hair samples of patients with breast cancer, whereas it was elevated in tumor tissues. CONCLUSIONS: There is a significant relationship between lowered serum Zn concentrations and risk of breast cancer onset or recurrences in women, but because of high heterogeneity, we recommend other primary studies.


Assuntos
Neoplasias da Mama/sangue , Zinco/sangue , Adulto , Mama/metabolismo , Feminino , Cabelo/química , Humanos , Pessoa de Meia-Idade , Viés de Publicação , Fatores de Risco , Sensibilidade e Especificidade
6.
Technol Cancer Res Treat ; 18: 1533033819870823, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31431135

RESUMO

Successful therapies for patients with breast cancer often lose their initial effectiveness. Thus, identifying new molecular targets is a constant goal in the fight against breast cancer. Gpn3 is a protein required for RNA polymerase II nuclear targeting in both yeast and human cells. We investigated here the effect of suppressing Gpn3 expression on cell proliferation in a progression series of isogenic cell lines derived from the nontumorigenic MCF-10A breast cells that recapitulate different stages of breast carcinogenesis. Gpn3 protein levels were comparable in all malignant derivatives of the nontumorigenic MCF-10A cells. shRNA-mediated inhibition of Gpn3 expression markedly decreased cell proliferation in all MCF-10A sublines. A fraction of the largest RNA polymerase II subunit Rpb1 was retained in the cytoplasm, but most Rpb1 remained nuclear after suppressing Gpn3 in all cell lines studied. Long-term proliferation experiments in cells with suppressed Gpn3 expression resulted in the eventual loss of all isogenic cell lines but MCF-10CA1d.cl1. In MCF-10CA1d.cl1 cells, Gpn3 knockdown reduced the proliferation of breast cancer stem cells as evaluated by mammosphere assays. After the identification that Gpn3 plays a key role in cell proliferation in mammary epithelial cells independent of the degree of transformation, we also analyzed the importance of Gpn3 in other human breast cancer cell lines from different subtypes. Gpn3 was also required for cell proliferation and nuclear translocation of RNA polymerase II in such cellular models. Altogether, our results show that Gpn3 is essential for breast cancer cell proliferation regardless of the transformation level, indicating that Gpn3 could be considered a molecular target for the development of new antiproliferative therapies. Importantly, our analysis of public data revealed that Gpn3 overexpression was associated with a significant decrease in overall survival in patients with estrogen receptor-positive and Human epidermal growth factor receptor 2 (HER2+) breast cancer, supporting our proposal that targeting Gpn3 could potentially benefit patients with breast cancer.


Assuntos
Neoplasias da Mama/genética , Carcinogênese/genética , GTP Fosfo-Hidrolases/genética , Receptor ErbB-2/genética , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/epidemiologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Intervalo Livre de Doença , Células Epiteliais , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia
7.
Medicine (Baltimore) ; 98(27): e16306, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31277170

RESUMO

This study investigated the effect of sex hormones on F-fluorodeoxyglucose (FDG) uptake in normal breast tissue.The retrospective study included 249 premenopausal women (median age, 45 years) who were diagnosed with unilateral breast cancer and underwent FDG positron emission tomography/computed tomography and hormone tests. The volume of interest was within the contralateral normal breast and the standardized uptake values (SUVs) were measured. The correlations of sex hormones (including estrogen, progesterone, testosterone, follicle-stimulating hormone [FSH] and luteinizing hormone [LH]) with the SUVs of the normal breast were analyzed.There was a weak negative correlation between age and breast FDG uptake (P = .012, Spearman coefficient = -.16 for the maximum standardized uptake values [SUVmax]), especially in the luteal phase group (P = .005, Spearman coefficient = -.27 for SUVmax). The SUVs of normal breast tissue were increased when progesterone levels were higher (P = .043, Spearman coefficient = .13 for SUVmax). In the irregular menstrual cycle group, FDG uptake in the breast decreased as FSH (P = .027, Spearman coefficient = -.42 for SUVmax) and LH (P = .048, Spearman coefficient = -.44 for SUVmax) increased.Glucose metabolism of normal breast tissue decreases with age, and progesterone weakly affects breast FDG uptake. Gonadotropins may affect breast FDG uptake in premenopausal women with irregular menstrual cycles.


Assuntos
Mama/metabolismo , Estradiol/sangue , Fluordesoxiglucose F18/farmacocinética , Hormônio Foliculoestimulante/sangue , Hormônio Luteinizante/sangue , Progesterona/sangue , Testosterona/sangue , Adulto , Mama/diagnóstico por imagem , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/metabolismo , Feminino , Humanos , Pessoa de Meia-Idade , Tomografia Computadorizada com Tomografia por Emissão de Pósitrons , Compostos Radiofarmacêuticos/farmacocinética , Valores de Referência , Estudos Retrospectivos
8.
Mar Drugs ; 17(8)2019 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-31349625

RESUMO

Breast cancer is the most common cancer type and a primary cause of cancer mortality among females worldwide. Here, we analyzed the anticancer efficacy of a novel bromochlorinated monoterpene, PPM1, a synthetic analogue of polyhalogenated monoterpenes from Plocamium red algae and structurally similar non-brominated monoterpenes. PPM1, but not the non-brominated monoterpenes, decreased selectively the viability of several triple-negative as well as triple-positive breast cancer cells with different p53 status without significantly affecting normal breast epithelial cells. PPM1 induced accumulation of triple-negative MDA-MB-231 cells with 4N DNA content characterized by decreased histone H3-S10/T3 phosphorylation indicating cell cycle arrest in the G2 phase. Western immunoblot analysis revealed that PPM1 treatment triggered an initial rapid activation of Aurora kinases A/B/C and p21Waf1/Cip1 accumulation, which was followed by accumulation of polyploid >4N cells. Flow cytometric analysis showed mitochondrial potential disruption, caspase 3/7 activation, phosphatidylserine externalization, reduction of the amount polyploid cells, and DNA fragmentation consistent with induction of apoptosis. Cell viability was partially restored by the pan-caspase inhibitor Z-VAD-FMK indicating caspase contribution. In vivo, PPM1 inhibited growth, proliferation, and induced apoptosis in MDA-MB-231 xenografted onto the chick chorioallantoic membrane. Hence, Plocamium polyhalogenated monoterpenes and synthetic analogues deserve further exploration as promising anticancer lead compounds.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias da Mama/tratamento farmacológico , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Monoterpenos/farmacologia , Antineoplásicos/farmacologia , Mama/efeitos dos fármacos , Mama/metabolismo , Neoplasias da Mama/metabolismo , Inibidores de Caspase/farmacologia , Caspases/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Fase G2/efeitos dos fármacos , Histonas/metabolismo , Humanos , Células MCF-7 , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Plocamium/química , Rodófitas/química
9.
Cancer Causes Control ; 30(10): 1103-1111, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31352658

RESUMO

PURPOSE: High mammographic breast density is a strong, well-established breast cancer risk factor. Whether stem cells may explain high breast cancer risk in dense breasts is unknown. We investigated the association between breast density and breast cancer risk by the status of stem cell markers CD44, CD24, and ALDH1A1 in the tumor. METHODS: We included 223 women with primary invasive or in situ breast cancer and 399 age-matched controls from Mayo Clinic Mammography Study. Percent breast density (PD), absolute dense area (DA), and non-dense area (NDA) were assessed using computer-assisted thresholding technique. Immunohistochemical analysis of the markers was performed on tumor tissue microarrays according to a standard protocol. We used polytomous logistic regression to quantify the associations of breast density measures with breast cancer risk across marker-defined tumor subtypes. RESULTS: Of the 223 cancers in the study, 182 were positive for CD44, 83 for CD24 and 52 for ALDH1A1. Associations of PD were not significantly different across t marker-defined subtypes (51% + vs. 11-25%: OR 2.83, 95% CI 1.49-5.37 for CD44+ vs. OR 1.87, 95% CI 0.47-7.51 for CD44-, p-heterogeneity = 0.66; OR 2.80, 95% CI 1.27-6.18 for CD24+ vs. OR 2.44, 95% CI 1.14-5.22 for CD24-, p-heterogeneity = 0.61; OR 3.04, 95% CI 1.14-8.10 for ALDH1A1+ vs. OR 2.57. 95% CI 1.30-5.08 for ALDH1A1-, p-heterogeneity = 0.94). Positive associations of DA and inverse associations of NDA with breast cancer risk were similar across marker-defined subtypes. CONCLUSIONS: We found no evidence of differential associations of breast density with breast cancer risk by the status of stem cell markers. Further studies in larger study populations are warranted to confirm these associations.


Assuntos
Biomarcadores Tumorais/metabolismo , Densidade da Mama , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/metabolismo , Mamografia , Idoso , Mama/diagnóstico por imagem , Mama/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , Pessoa de Meia-Idade , Fatores de Risco , Células-Tronco/metabolismo
10.
Nat Commun ; 10(1): 2725, 2019 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-31221963

RESUMO

Recent research has shown that many types of cancers take control of specific metabolic processes. We compiled metabolic networks corresponding to four healthy and cancer tissues, and analysed the healthy-cancer transition from the metabolic flux change perspective. We used a Probabilistic Minimum Dominating Set (PMDS) model, which identifies a minimum set of nodes that act as driver nodes and control the entire network. The combination of control theory with flux correlation analysis shows that flux correlations substantially increase in cancer states of breast, kidney and urothelial tissues, but not in lung. No change in the network topology between healthy and cancer networks was observed, but PMDS analysis shows that cancer states require fewer controllers than their corresponding healthy states. These results indicate that cancer metabolism is characterised by more streamlined flux distributions, which may be focused towards a reduced set of objectives and controlled by fewer regulatory elements.


Assuntos
Redes e Vias Metabólicas/fisiologia , Modelos Biológicos , Modelos Estatísticos , Neoplasias/metabolismo , Algoritmos , Mama/metabolismo , Simulação por Computador , Humanos , Rim/metabolismo , Pulmão/metabolismo , Análise do Fluxo Metabólico , Neoplasias/patologia , Urotélio/metabolismo
11.
Life Sci ; 232: 116610, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31254584

RESUMO

AIMS: The aim of this study was the characterization of the in vitro cytotoxic properties of a recently isolated diterpene compound, 7ß-acetoxy-20-hydroxy-19,20-epoxyroyleanone (compound 1), extracted from Salvia corrugata, versus human cell lines. MAIN METHODS: We used as model study immortalized breast epithelial cells MCF10A and two ERBB2+ breast cancer (BCa) cell lines, SKBR-3 and BT474. Compound 1 was isolated by methanolic extraction from regenerated shoots of Salvia corrugata Vahl, and purified by high pressure liquid chromatography (HPLC). Flow cytometry (FCM) was employed for cell cycle, apoptosis and reactive oxygen species (ROS) analysis. Cell morphology was assessed by immunofluorescence and transmission electron microscopy (TEM). KEY FINDINGS: Compound 1 inhibited cell survival of all breast cell lines. In particular, compound 1 promoted cell cycle arrest in the G0/G1 phase and apoptosis along with impairment of the mitochondrial function, which was reflected in a gross alteration of the mitochondrial network structure. Furthermore, we also detected a potent activation of the ERK1/2 kinase, which suggested the induction of reactive oxygen species (ROS). Partial rescue of survival obtained with n-acetylcysteine (NAC) when coadminstered with compound 1 further supported a contribution of ROS mediated mechanisms to the growth-arrest and proapoptotic activity of compound 1 in both BCa cell lines. ROS production was indeed confirmed in SKBR-3. SIGNIFICANCE: Our findings show that compound 1 has a cytotoxic activity against both human normal and cancer cell lines derived from breast epithelia, which is mediated by ROS generation and mitochondrial damage.


Assuntos
Mama/efeitos dos fármacos , Diterpenos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Células Epiteliais/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Mama/citologia , Mama/metabolismo , Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Diterpenos/isolamento & purificação , Células Epiteliais/metabolismo , Feminino , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fase de Repouso do Ciclo Celular/efeitos dos fármacos
12.
Endocrinology ; 160(9): 2074-2084, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31150047

RESUMO

Stress decreases milk components such as milk protein and milk yield. The objective of this study was to investigate whether noradrenaline (NA) in milk constituted a factor associated with stress-induced changes in milk proteins such as ß-casein. Breast milk obtained from eight healthy, nursing women contained NA at concentrations ranging from 12.7 to 115.5 nM. The expression of tyrosine hydroxylase (TH), a rate-limiting enzyme of NA synthesis, was observed in primary normal human mammary epithelial cells (HMECs), and in MCF-12A and MCF-10A cell lines. The mean NA concentration in culture medium used by MCF-12A transfected with TH small interfering RNA (siRNA) was significantly lower than that of cells transfected with control siRNA. NA concentration in milk in restraint-stressed nursing mice was significantly higher than that in nonstressed nursing mice, owing to elevated TH expression in the mammary epithelium. The mean ß-casein concentration in milk in restraint-stressed mice was significantly lower than that in nonstressed mice. NA treatment resulted in a concentration-dependent decrease in ß-casein expression in HMECs. ß2 adrenergic receptor (ADRB2) expression was observed in HMECs, MCF-12A, and MCF-10A, and immunohistochemical analysis of ADRB2 using mammary epithelium sections obtained from mice at day 10 of lactation showed that ADRB2 was expressed at the apical membrane of mammary epithelium. Treatment with salbutamol, an ADRB2 stimulant, decreased ß-casein expression in a concentration-dependent manner in MCF-12A. Our results showed that endogenous NA derived from mammary epithelial cells likely comprises one of the factors involved in stress-induced changes in milk proteins such as ß-casein.


Assuntos
Caseínas/análise , Leite/química , Norepinefrina/fisiologia , Estresse Psicológico/metabolismo , Adulto , Animais , Mama/metabolismo , Células Cultivadas , Feminino , Humanos , Camundongos , Gravidez , Receptores Adrenérgicos beta 2/análise , Tirosina 3-Mono-Oxigenase/genética
13.
Cells ; 8(6)2019 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-31216647

RESUMO

Human breast cancer is characterized by a high degree of inter-patients heterogeneity in terms of histology, genomic alterations, gene expression patterns, and metastatic behavior, which deeply influences individual prognosis and treatment response. The main cause of mortality in breast cancer is the therapy-resistant metastatic disease, which sets the priority for novel treatment strategies for these patients. In the present study, we demonstrate that Patient Derived Xenografts (PDXs) that were obtained from metastatic and therapy-resistant breast cancer samples recapitulate the wide spectrum of the disease in terms of histologic subtypes and mutational profiles, as evaluated by whole exome sequencing. We have integrated genomic and transcriptomic data to identify oncogenic and actionable pathways in each PDX. By taking advantage of primary short-term in vitro cultures from PDX tumors, we showed their resistance to standard chemotherapy (Paclitaxel), as seen in the patients. Moreover, we selected targeting drugs and analyzed PDX sensitivity to single agents or to combination of targeted and standard therapy on the basis of PDX-specific genomic or transcriptomic alterations. Our data demonstrate that PDXs represent a suitable model to test new targeting drugs or drug combinations and to prioritize personalized therapeutic regimens for pre-clinal and clinical tests.


Assuntos
Neoplasias da Mama/genética , Resistencia a Medicamentos Antineoplásicos/genética , Medicina de Precisão/métodos , Animais , Mama/metabolismo , Modelos Animais de Doenças , Feminino , Xenoenxertos/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos NOD , Metástase Neoplásica/genética , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
14.
Zhonghua Zhong Liu Za Zhi ; 41(5): 331-337, 2019 May 23.
Artigo em Chinês | MEDLINE | ID: mdl-31137165

RESUMO

Objective: To investigate the differential expression profiles of circular RNA (circRNA) in human epidermal growth factor receptor 2 (HER-2) positive breast cancer cells and normal mammary epithelial cells, and to develop novel diagnostic and therapeutic markers for HER-2 positive breast cancer. Methods: Total RNA were extracted from HER-2 positive breast cancer cell SK-BR-3 and normal mammary epithelial cell MCF10A. RNA quality was detected using NanoDrop ND-1000. Rnase R was applied to remove linear RNA and enrich circRNAs. After amplification and reverse transcription into fluorescent complementary RNA (cRNA) using random primer, the labeled cRNAs were hybridized onto the Arraystar Human circRNA Arrays. The raw data were extracted and the acquired array images were subjected to quantile normalization followed by heat map and volcano plot analysis. The expression of circRNAs with large fold change was verified by real-time quantitative polymerase chain reaction (RT-qPCR). Finally, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed in the differentially expressed circRNAs and circRNA-microRNA (miRNA) network was constructed. Results: The total RNA extracted from SK-BR-3 and MCF10A had high integrity and quality. The expression profiles of circRNA in SK-BR-3 and MCF10A cells were significantly different shown by fluorescent expression signals. Compared with MCF10A cells, there were 6 584 up-regulated circRNAs and 6254 down-regulated circRNAs in SK-BR-3 cells. There were 348 circRNAs with |log(2)FC|≥2, of which 153 were up-regulated and 195 were down-regulated. Moreover, 8 circRNAs with |log(2)FC|>5. Among them, 5 were up-regulated in SK-BR-3 cells, including hsa_circRNA_074595 (|log(2)FC|=7.84), hsa_circRNA_074598 (|log(2)FC|=6.50), hsa_circRNA_085362 (|log(2)FC|=5.86), hsa_circRNA_101379 (|log(2)FC|=5.71) and hsa_circRNA_406683 (|log(2)FC|=5.34); as well as 3 were down-regulated, including hsa_circRNA_021714 (|log(2)FC|=5.46), hsa_circRNA_100777 (|log(2)FC|=5.40), and hsa_circRNA_100796 (|log(2)FC|=5.03). The expression levels of hsa_circRNA_074595, hsa_circRNA_074598 and hsa_circRNA_100777 were further validated by RT-qPCR in consistent with the results of microarray. GO analysis showed that differentially expressed circRNAs were significantly enriched in the biological process of heart development (P<0.001), cellular component in the cell adhesion-related components (P<0.001), molecular function in protein serine/threonine kinase activity (P<0.001). KEGG analysis revealed that differentially expressed circRNAs were significantly enriched in the PI3K-Akt signaling pathway. Conclusions: The expression profile of circRNA in HER-2 positive breast cancer cells is significantly different from that in normal mammary epithelial cells. The differentially expressed circRNAs may be served as potential diagnostic or therapeutic targets for HER-2 positive breast cancer.


Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Mama/metabolismo , Células Epiteliais/metabolismo , RNA não Traduzido/biossíntese , Receptor ErbB-2/metabolismo , Mama/citologia , Células Epiteliais/citologia , Feminino , Perfilação da Expressão Gênica , Humanos , MicroRNAs/biossíntese , MicroRNAs/genética , RNA não Traduzido/genética , Receptor ErbB-2/genética
15.
Radiol Phys Technol ; 12(3): 260-267, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31129787

RESUMO

In this study, we aimed to develop a hybrid method for automated detection of high-uptake regions in the breast and axilla using dedicated breast positron-emission tomography (db PET) and whole-body PET/computed tomography (CT) images. In our proposed method, high-uptake regions in the breast and axilla were detected using db PET images and whole-body PET/CT images. In db PET images, high-uptake regions in the breast were detected using adaptive thresholding technique based on the noise characteristics. In whole-body PET/CT images, the region of the breast that includes the axilla was first extracted using CT images. Next, high-uptake regions in the extracted breast region were detected on the PET images. By integration of the results of the two types of PET images, a final candidate region was obtained. In the experiments, the accuracy of extracting the region of the breast and detection ability was evaluated using clinical data. As a result, all breast regions were extracted correctly. The sensitivity of detection was 0.765, and the number of false positive cases were 1.8, which was 30% better than those on whole-body PET/CT alone. These results suggested that the proposed method, combining the two types of PET images is effective for improving detection performance.


Assuntos
Mama/diagnóstico por imagem , Mama/metabolismo , Processamento de Imagem Assistida por Computador/métodos , Tomografia Computadorizada com Tomografia por Emissão de Pósitrons , Imagem Corporal Total , Automação , Transporte Biológico , Humanos
16.
J BUON ; 24(2): 535-542, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31128002

RESUMO

PURPOSE: This study aimed to investigate the correlation of long non-coding RNA (lncRNA) TCONS_l2_00002973 expression with clinicopathological features as well as overall survival (OS) in triple-negative breast cancer (TNBC) patients, and further explore the regulatory effect of lncRNA TCONS_l2_00002973 on cells' proliferation and apoptosis in TNBC cells. METHODS: 96 TNBC patients undergoing surgery were consecutively enrolled in this prospective cohort study. LncRNA TCONS_l2_00002973 expression in tumor tissue sample and adjacent non-tumor sample was detected by qPCR. Normal control (NC) shRNA (NC(-) group), lncRNA TCONS_l2_00002973 shRNA (Lnc(-) group), NC overexpression (NC(+) group) and lncRNA TCONS_l2_00002973 overexpression plasmids (Lnc(+) group) were transfected into MDA-MB-231 breast cancer cells. Cell proliferation ability, cell apoptosis rate and apoptosis-related protein expressions were detected using CCK-8 assay, annexin V (AV)/propidium iodide (PI) assay and Western blot assay respectively. RESULTS: LncRNA TCONS_l2_00002973 expression was lower in tumor tissue compared to paired adjacent tissue (p<0.001), and its low expression was associated with increased T stage (p=0.002), raised N stage (p=0.003) and advanced TNM stage (p=0.005) in TNBC patients. Poor OS was found in lncRNA TCONS_l2_00002973 low expression group compared to lncRNA TCONS_l2_00002973 high expression group (p=0.006). Further in vitro experiments disclosed that lncRNA TCONS_l2_00002973 expression was reduced in various breast cancer cell lines compared to normal breast cell line, and lncRNA TCONS_l2_00002973 repressed cell proliferation and enhanced cells apoptosis in MDA-MB-231 cells. CONCLUSIONS: LncRNA TCONS_l2_00002973 correlates with less advanced tumor stage and favorable survival, and it also inhibits cancer cells proliferation while enhances apoptosis in TNBC.


Assuntos
Proliferação de Células/genética , RNA Longo não Codificante/genética , Neoplasias de Mama Triplo Negativas/genética , Apoptose/genética , Biomarcadores Tumorais/genética , Mama/metabolismo , Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Intervalo Livre de Doença , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Estimativa de Kaplan-Meier , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neoplasias de Mama Triplo Negativas/patologia
17.
Anticancer Res ; 39(5): 2647-2659, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31092464

RESUMO

BACKGROUND/AIM: The aim of the present study was to analyze metastasized breast cancer (BC) patients with regard to the discordance of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). We especially aimed to analyze the association between the change of tumor biology and previous treatment or metastatic sites. PATIENTS AND METHODS: Patients with metastasized BC who were treated at the Department of Gynecology/Breast Center of the University Hospital of Cologne were analyzed. RESULTS: Loss of HER2 occurred more frequently in lymph node metastases that were not in the axillary region (p=0.026). Letrozole showed a significant correlation with loss of ER and/or PR (p=0.041). Improved overall survival and post-metastasis survival were noticed with a gain of HER2 (p=0.044 and p=0.009, respectively) and concordant positive ER and PR status (p=0.002 and p=0.001, respectively). CONCLUSION: The discordance of receptors and the dependence of BC on therapies as well as metastatic sites stresses the necessity of early sample taking to offer patients suitable therapy options.


Assuntos
Biomarcadores Tumorais/genética , Neoplasias da Mama/genética , Recidiva Local de Neoplasia/genética , Segunda Neoplasia Primária/genética , Adulto , Idoso , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Receptor alfa de Estrogênio/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Linfonodos/metabolismo , Linfonodos/patologia , Pessoa de Meia-Idade , Metástase Neoplásica , Recidiva Local de Neoplasia/tratamento farmacológico , Recidiva Local de Neoplasia/patologia , Segunda Neoplasia Primária/tratamento farmacológico , Segunda Neoplasia Primária/patologia , Receptor ErbB-2/genética , Receptores de Progesterona/genética
18.
Nucleic Acids Res ; 47(10): 5086-5099, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-30982901

RESUMO

BRCA1-associated basal-like breast cancer originates from luminal progenitor cells. Breast epithelial cells from cancer-free BRCA1 mutation carriers are defective in luminal differentiation. However, how BRCA1 deficiency leads to lineage-specific differentiation defect is not clear. BRCA1 is implicated in resolving R-loops, DNA-RNA hybrid structures associated with genome instability and transcriptional regulation. We recently showed that R-loops are preferentially accumulated in breast luminal epithelial cells of BRCA1 mutation carriers. Here, we interrogate the impact of a BRCA1 mutation-associated R-loop located in a putative transcriptional enhancer upstream of the ERα-encoding ESR1 gene. Genetic ablation confirms the relevance of this R-loop-containing region to enhancer-promoter interactions and transcriptional activation of the corresponding neighboring genes, including ESR1, CCDC170 and RMND1. BRCA1 knockdown in ERα+ luminal breast cancer cells increases intensity of this R-loop and reduces transcription of its neighboring genes. The deleterious effect of BRCA1 depletion on transcription is mitigated by ectopic expression of R-loop-removing RNase H1. Furthermore, RNase H1 overexpression in primary breast cells from BRCA1 mutation carriers results in a shift from luminal progenitor cells to mature luminal cells. Our findings suggest that BRCA1-dependent R-loop mitigation contributes to luminal cell-specific transcription and differentiation, which could in turn suppress BRCA1-associated tumorigenesis.


Assuntos
Proteína BRCA1/genética , Mama/metabolismo , Elementos Facilitadores Genéticos , Células Epiteliais/metabolismo , Regulação Neoplásica da Expressão Gênica , Proteína BRCA1/metabolismo , Sistemas CRISPR-Cas , Carcinogênese , Diferenciação Celular , Receptor alfa de Estrogênio/genética , Feminino , Deleção de Genes , Genes BRCA1 , Células HEK293 , Heterozigoto , Humanos , Células MCF-7 , Mutação , Transcrição Genética
19.
BMC Cancer ; 19(1): 388, 2019 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31023265

RESUMO

BACKGROUND: Gene expression profiling provides key information for prognosis of breast cancer to establish treatment strategy. However, the genetic assessment should be available before induction of treatment to be useful for clinical practice. To evaluate the reliability of using needle biopsy samples for gene assays, we compared gene-expression profiling results between core needle biopsy (CNB) samples and surgical specimens in breast cancer. METHODS: Thirty-one paired, formalin-fixed, paraffin-embedded CNB and surgical specimen samples were selected from patients with hormone receptor-positive breast cancer. Total RNA was extracted from the samples and the risk classifications based on GenesWell BCT scores were compared. RESULTS: The BCT scores correlated between CNB samples and surgical specimens of hormone receptor-positive breast cancer (Pearson r = 0.66). The overall concordance rate of risk classification (high/low risk) was 83.9%. However, when the breast cancer does not contain intratumoral microcalcification, the concordance rate increased as 92.0%. And, when the breast cancer formed a solitary nodule (non-multifocal), the concordance rate increased up to 95.8%. CONCLUSION: Risk classification using the GenesWell BCT multigene kit with CNB samples could be considered reliable, when the breast cancer is a solitary nodule without intratumoral microcalcification. Such genetic profiling results should be helpful for establishing a treatment plan for hormone receptor-positive breast cancer before treatment induction.


Assuntos
Neoplasias da Mama/epidemiologia , Neoplasias da Mama/genética , Mama/metabolismo , Medição de Risco , Biópsia com Agulha de Grande Calibre , Mama/patologia , Neoplasias da Mama/patologia , Feminino , Humanos , Pessoa de Meia-Idade , Família Multigênica/genética , RNA , Receptor ErbB-2/genética , Receptores Estrogênicos/genética , Receptores de Progesterona/genética
20.
J Exp Clin Cancer Res ; 38(1): 173, 2019 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31023337

RESUMO

BACKGROUND: Breast cancer angiogenesis is key for metastasis and predicts a poor prognosis. Angiotensin-converting enzyme 2 (ACE2), as a member of the renin-angiotensin system (RAS), was reported to restrain the progression of hepatocellular carcinoma (HCC) and non-small cell lung cancer (NSCLC) through inhibiting angiogenesis. However, the relationship between ACE2 and breast cancer angiogenesis remains unclear. METHODS: The prognosis and relative gene selection were analysed using the GEPIA, GEO, TCGA and STRING databases. ACE2 expression in breast cancer tissue was estimated by reverse transcription-quantitative polymerase chain reaction (qPCR). Breast cancer cell migration, proliferation and angiogenesis were assessed by Transwell migration, proliferation, tube formation, and wound healing assays. The expression of vascular endothelial growth factor A (VEGFa) was detected by qPCR and Western blotting. The phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2), mitogen-activated protein kinase 1/2 (MEK1/2), and extracellular signal-regulated protein kinase 1/2 (ERK1/2) was examined by Western blotting. Breast cancer metastasis and angiogenesis in vivo were measured using a zebrafish model. RESULTS: ACE2 was downregulated in breast cancer patients. Patients with higher ACE2 expression had longer relapse-free survival (RFS). In vitro, ACE2 inhibited breast cancer migration. Meanwhile, ACE2 in breast cancer cells inhibited human umbilical vascular endothelial cell (HUVEC) proliferation, tube formation and migration. In the zebrafish model, ACE2 inhibited breast cancer cell metastasis, as demonstrated by analyses of the number of disseminated foci and the metastatic distance. Neo-angiogenesis was also decreased by ACE2. ACE2 downregulated the expression of VEGFa in breast cancer cells. Furthermore, ACE2 in breast cancer cells inactivated the phosphorylation of VEGFR2, MEK1/2, and ERK1/2 in HUVECs. CONCLUSIONS: Our findings suggest that ACE2, as a potential resister to breast cancer, might inhibit breast cancer angiogenesis through the VEGFa/VEGFR2/ERK pathway. TRIAL REGISTRATION: Retrospectively registered.


Assuntos
Neoplasias da Mama/genética , Neovascularização Patológica/genética , Peptidil Dipeptidase A/genética , Fator A de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Animais , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/patologia , Movimento Celular/genética , Proliferação de Células/genética , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Células Endoteliais da Veia Umbilical Humana , Humanos , Sistema de Sinalização das MAP Quinases/genética , Células MCF-7 , Metástase Neoplásica , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/patologia , Fosforilação , Peixe-Zebra
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