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1.
Food Chem ; 367: 130617, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34352696

RESUMO

The abuse application of glyphosate can result in a potential hazard for environment and human, however its ultrasensitive detection remains challenging. Herein, a Cu2+ modulated DNA-templated silver nanoclusters (DNA-AgNCs) sensor was constructed to sensitively determine glyphosate based on the turn-on fluorescence strategy. The fluorescence quenching of DNA-AgNCs occurred with the existence of Cu2+. Upon the presence of glyphosate, the functional groups on the surface of glyphosate could chelate with Cu2+, following the fluorescence recovery of DNA-AgNCs. Through the stoichiometric methods, we unveil that Cu2+-trigged fluorescence quenching mode is a combination of static and dynamic quenching with the static mode being predominant. In DNA-AgNCs/Cu2+ system, the carboxylate, amine, and phosphonate groups of glyphosate interact with Cu2+ through chelation, in which the carboxylate oxygen, the phosphonate oxygen atoms, and the monoprotonated secondary amine nitrogen atom and Cu2+ form chelate rings. This fluorescence sensor showed a desired linearity of glyphosate analysis under the optimum conditions, ranging from 15 to 100 µg/L with a low detection down to 5 µg/L. Moreover, the proposed sensor was successfully utilized to measure glyphosate in real samples, indicating a promising application in pesticide residues detection.


Assuntos
Nanopartículas Metálicas , Prata , DNA/genética , Glicina/análogos & derivados , Humanos
2.
J Biomed Nanotechnol ; 17(8): 1525-1534, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-34544530

RESUMO

The development of science and technology has deepened people's understanding of cancer, changing the management of malignant tumors in the medical field. Given the common precancerous characteristics of colorectal cancer (CRC), researchers studied early CRC screening. The complexity of traditional diagnostics forced medical staff to speed up CRC innovation early screening methods. Here, we prepared nano-colloidal gold raw materials with different particle sizes (15 and 30 nm) and observed the morphological characteristics and properties of the materials. Simultaneously, the nanocolloidal gold double antibody sandwich kit was designed through the optimum pH value and protein content screening experiment. The results of clinical enteroscopy confirmed the important guiding significance of the equipment in early CRC screening.


Assuntos
Neoplasias Colorretais , Sangue Oculto , Cromatografia de Afinidade , Neoplasias Colorretais/diagnóstico , Detecção Precoce de Câncer , Coloide de Ouro , Humanos
3.
Reprod Biol Endocrinol ; 19(1): 101, 2021 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-34215268

RESUMO

BACKGROUND: Human Ectopic Pregnancy (hEP) is the second most common cause of pregnancy-related deaths in the first trimester. Without timely detection, EPs can lead to an increased rate of infertility and an elevated risk for future tubal EPs. In addition, most studies in the field focus on the effect of the fallopian tube (maternal factors) and ignore epigenetic changes in genes and proteins of the embryo, which may also cause EPs. Therefore, the present study hypothesized that embryos also play an important role in the development of EP. The study also speculated that DNA methylation is associated with ectopic pregnancy. Consequently, the effects of DNA methylation on the occurrence and development of ectopic pregnancy were investigated. Moreover, genome-wide DNA methylation of chorionic tissue from ectopic and intrauterine pregnancies was detected using Illumina HumanMethylation450 arrays. RESULTS: Forty-three hypermethylated genes involved in the regulation of adhesion as well as gene transcription and translation were identified. Furthermore, the PPI network showed that AMOTL1, SDR42E1, CAMTA1, PIP5K1C, KIAA1614, TSTD1 and DNER may play important roles in the occurrence and development of ectopic pregnancy. In addition, SDR42E1, CAMTA1 and TSTD1 displayed higher levels of methylation in ectopic pregnancy while PIP5K1C and DNER showed low degrees of methylation. CONCLUSIONS: The study reveals that abnormal increase in methylation may be an early indicator or an inducer of ectopic pregnancy. In addition, AMOTL1, SDR42E1, CAMTA1, PIP5K1C, KIAA1614, TSTD1 and DNER might play important roles in the occurrence and development of ectopic pregnancy. However, the specific molecular mechanisms are still unclear and require further studies.

4.
Aging (Albany NY) ; 13(10): 13954-13967, 2021 05 12.
Artigo em Inglês | MEDLINE | ID: mdl-33982673

RESUMO

To examine the role of S100B in genetic susceptibility to Alzheimer's disease (AD), we conducted a case-control study to analyze four polymorphism loci (rs2839364, rs1051169, rs2300403, and rs9722) of the S100B gene and AD risk. We found an independent increased risk of AD in ApoE ε4(-) subjects carrying the rs9722 AA-genotype (OR = 2.622, 95% CI = 1.399-4.915, P = 0.003). Further investigation revealed the serum S100B levels to be lower in rs9722 GG carriers than in rs9722 AA carriers (P = 0.003). We identified three miRNAs (miR-340-3p, miR-593-3p, miR-6827-3p) in which the seed match region covered locus rs9722. Luciferase assays indicated that the rs9722 G allele has a higher binding affinity to miR-6827-3p than the rs9722 A allele, leading to a significantly decreased fluorescence intensity. Subsequent western blot analysis showed that the S100B protein level of SH-SY5Y cells, which carry the rs9722 G allele, decreased significantly following miR-6827-3p stimulation (P = 0.009). The present study suggests that the rs9722 polymorphism may upregulate the expression of S100B by altering the miRNA binding capacity and may thus increase the AD risk. This finding would be of great help for the early diagnosis of AD.


Assuntos
Doença de Alzheimer/sangue , Doença de Alzheimer/genética , Predisposição Genética para Doença , MicroRNAs/metabolismo , Polimorfismo de Nucleotídeo Único/genética , Subunidade beta da Proteína Ligante de Cálcio S100/sangue , Subunidade beta da Proteína Ligante de Cálcio S100/genética , Idoso , Sequência de Bases , Estudos de Casos e Controles , Linhagem Celular Tumoral , Ensaios Enzimáticos , Feminino , Frequência do Gene/genética , Haplótipos/genética , Humanos , Luciferases/metabolismo , Masculino , MicroRNAs/genética , Ligação Proteica/genética
5.
J Assist Reprod Genet ; 37(9): 2053-2079, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32562095

RESUMO

PURPOSE: Patients with Klinefelter syndrome (KS) who receive assisted reproductive technology (ART) treatment often experience poor pregnancy rates due to decreased fertilization, cleavage, and implantation rates and even an increased miscarriage rate. Mounting evidence from recent studies has shown that various technological advances and approaches could facilitate the success of ART treatment for KS patients. In this review, we summarize the methods for guiding KS patients during ART and for developing optimal strategies for preserving fertility, improving pregnancy rate and live birth rate, and avoiding the birth of KS infants. METHODS: We searched PubMed and Google Scholar publications related to KS patients on topics of controlled ovarian stimulation protocols, sperm extraction, fertility preservation, gamete artificial activation, round spermatid injection (ROSI), and non-invasive prenatal screening (PGD) methods. RESULTS: This review outlines the different ovulation-inducing treatments for female partners according to the individual sperm status in the KS patient. We further summarize the methods of retrieving sperm, storing, and freezing rare sperm. We reviewed different methods of gamete artificial activation and discussed the feasibility of ROSI for sterile KS patients who absolutely lack sperm. The activation of eggs in the process of intracytoplasmic sperm injection and non-invasive PGD are urgently needed to prevent the birth of KS infants. CONCLUSION: The integrated strategies will pave the way for the establishment of ART treatment approaches and improve the clinical outcome for KS patients.


Assuntos
Implantação do Embrião/genética , Síndrome de Klinefelter/terapia , Técnicas de Reprodução Assistida/tendências , Coeficiente de Natalidade , Feminino , Preservação da Fertilidade/tendências , Humanos , Síndrome de Klinefelter/genética , Síndrome de Klinefelter/patologia , Masculino , Gravidez , Taxa de Gravidez , Injeções de Esperma Intracitoplásmicas/tendências
6.
Mikrochim Acta ; 187(6): 341, 2020 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-32444888

RESUMO

N-Doped silicon quantum dots (N-SiQD) were synthesized using N-[3-(trimethoxysily)propyl]-ethylenediamine and citric acid as silicon source and reduction agent, respectively. The N-SiQD shows a strong blue fluorescence with a high quantum yield of about 53%. It is found that a selective static quenching process occurs between N-SiQDs and Cu2+. Glyphosate can inhibit this phenomenon and trigger the rapid fluorescence enhancement of the quenched N-SiQDs/Cu2+ system due to the specific interaction between Cu2+ and glyphosate. With such a design, a turn-on fluorescent nanoprobe based on N-SiQD/Cu2+ system was established for rapid determination of glyphosate. The determination signal of N-SiQD/Cu2+ was measured at the optimum emission wavelength of 460 nm after excitation at 360 nm. Under optimal conditions, the turn-on nanoprobe showed a linear relationship between fluorescent response and glyphosate concentrations in the range 0.1 to 1 µg mL-1. The limit of determination was calculated to 7.8 ng mL-1 (3σ/S). Satisfactory recoveries were obtained in the determination of spiked water samples, indicating the potential use for environmental monitoring. Graphical abstract Schematic representation of N-SiQD/Cu2+ system for glyphosate determination. Fluorescence quenching of N-SiQDs induced by copper ions and the succedent fluorescent "turn on" triggered by glyphosate.


Assuntos
Corantes Fluorescentes/química , Glicina/análogos & derivados , Pontos Quânticos/química , Cobre/química , Fluorescência , Glicina/análise , Lagos/análise , Nitrogênio/química , Rios/química , Silício/química , Espectrometria de Fluorescência/métodos , Poluentes Químicos da Água/análise
7.
Mikrochim Acta ; 186(4): 259, 2019 03 28.
Artigo em Inglês | MEDLINE | ID: mdl-30923924

RESUMO

A fluorometric method was developed for the determination of the insecticide cartap. It is based on the use of green emitting carbon dots (CDs) and gold nanoparticles (Au NPs). The CDs were prepared from phenol and ethylene diamine by a hydrothermal route. They have excitation/emission maxima at 410/513 nm) and a fluorescence quantum yield of 29%. They were characterized by TEM, Raman, XRD, XPS, FT-IR, UV and fluorescence spectroscopies. The green fluorescence of the CDs is strongly reduced by the red-colored Au NPs because of an inner filter effect. Upon addition of cartap, it will cause the aggregation of the Au NPs owing to Au-N interaction between Au NPs and cartap to form purple colored aggregates with spectra that do not overlap the green emission of the CDs. Hence, their fluorescence is restored. Under optimum conditions, the method allows for the quantitation of cartap in the 5-300 nM concentration range, and the detection limit is 3.8 nM. The method was successfully applied to the determination of cartap in spiked real samples and gave satisfactory results. Graphical abstract Schematic presentation of green emitting carbon dots for sensitive fluorometric determination of cartap based on its aggregation effect on gold nanoparticles.

8.
Am J Transl Res ; 11(1): 269-279, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30787985

RESUMO

Gestational diabetemellitus (GDM) is a condition whereby a mother's glucose tolerance is impaired with onset or first recognition during pregnancy which is not either type 1 or type 2 diabetes mellitus. Oxidative stress plays an essential role in diabetes, however, whether it also includes in GDM has not been fully clarified. Therefore, we investigated the changes of oxidative stress biomarkers and their relationship with pregnancy outcomes in patients with GDM. The serum and placenta were collected for absorbance-based assay and immunohistochemistry assay (IHC). The patients' clinical information was collected and the pregnancy outcome was tracked. It was found that elder age is a risk factor to result in GDM. Moreover, GDM patients showed poor clinical factors or outcomes including higher prepregnancy weight and BMI value, premature delivery, higher rates of cesarean delivery, macrosomia, premature rupture of fetal membranes (PROM). Increasing serum MDA level and decline GSH and SOD levels were observed in GDM patients. Meanwhile, HO-1, Nrf2 and NQO1 overexpressed in GDM placental tissues. In the GDM group, MDA level was negatively associated with prepregnancy weight, while, SOD level was positively correlated with neonatal birth weight. We found an intensive relationship between SOD content and preterm birth in the GDM group. There is no significant difference between the level of MDA/GSH and neonatal birth weight as well as preterm birth. MDA, GSH and SOD levels were not associated with an increased risk of cesarean delivery or PROM. This study indicates aberrant expression of oxidative stress related proteins affects the pregnancy outcome of GDM patients.

9.
Anal Sci ; 35(4): 441-448, 2019 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-30606912

RESUMO

In this paper, an ultrasensitive electrochemical biosensor based on carboxylated multi-walled carbon nanotube/molybdenum disulfide composites (MWCNTs-COOH/MoS2) for the detection of KRAS gene is described. An easy, low-cost method, named one-step hydrothermal, was used for the synthesize of MWCNTs-COOH/MoS2 nanocomposites, and scanning electronic microscopy (SEM), high resolution transmission electron microscopy (HRTEM), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD) were used for characterizing the prepared composites. Furthermore, cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were employed for an electrochemical performance study of this biosensor. Under optimal conditions, the detection limit of target DNA achieved down to 3 fM (S/N = 3) with high sensitivity; the linear range with the logarithm of the concentrations of target DNA varied from 1.0 × 10-14 to 1.0 × 10-7 M. Finally, the practicality of our proposed sensor was verified by a determination of the KRAS gene in human serum samples with good accuracy and high precision due to the excellent conductivity and large active surface area of the MWCNTs-COOH/MoS2 nanocomposites. This proposed biosensor thus provides a practical method for the rapid and sensitive analysis of gene detection.


Assuntos
Técnicas Biossensoriais , Ácidos Carboxílicos/química , DNA/química , Dissulfetos/química , Técnicas Eletroquímicas , Molibdênio/química , Nanotubos de Carbono/química , Proteínas Proto-Oncogênicas p21(ras)/genética , Humanos , Tamanho da Partícula , Proteínas Proto-Oncogênicas p21(ras)/sangue , Propriedades de Superfície
10.
Front Genet ; 9: 540, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30555509

RESUMO

To determine the role of A disintegrin and metalloproteinase 10 (ADAM10) in genetic susceptibility to Alzheimer's disease (AD) in a representative Chinese sample, we genotyped 362 AD patients and 370 healthy controls for the rs514049A/C and rs653765C/T polymorphisms in the ADAM10 promoter using the SNaPshot technique. We also examined the potential impact of these polymorphisms on the plasma level of soluble receptor for advanced glycation end products (sRAGE), a decoy receptor whose reduction has been associated with a higher risk of AD. Additionally, a meta-analysis was performed using the present study and the largest GWAS from the International Genomics of Alzheimer's Project (IGAP). No significant differences were found in the distributions of genotypes or alleles between AD patients and control subjects. However, age-at-onset stratification analysis revealed that there were significant differences in the genotypes (P = 0.015) and alleles (P = 0.006) of the rs653765 SNP. Furthermore, patients with the rs653765 CC genotype showed a lower ADAM10 level and a faster cognitive deterioration than those in patients with the CT/TT genotype in late-onset AD (LOAD), and the rs653765 CC polymorphism was able to regulate the production of the ADAM10 substrate sRAGE. In contrast, the rs514049 polymorphism was not statistically associated with AD. In the meta-analysis, we observed that both rs514049 (A allele vs. C allele, P = 0.002) and rs653765 (C allele vs. T allele, P = 0.004) were associated with AD risk. The present study indicated that the rs653765 polymorphism might be associated with the risk and development of LOAD; in particular, the risk genotype, CC, may decrease the expression of ADAM10, influencing the plasma levels of sRAGE, and thus may be correlated with the clinical progression of AD.

11.
J Alzheimers Dis ; 66(3): 887-899, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30400091

RESUMO

Alzheimer's disease (AD) is an insidious and progressive neurodegenerative disease. The main pathological features of AD are the formation of amyloid-ß deposits in the anterior cerebral cortex and hippocampus as well as the formation of intracellular neurofibrillary tangles. Thus far, accumulating evidence shows that glycation is closely related to AD. As a final product resulting from the crosslinking of a reducing sugar or other reactive carbonyls and a protein, the advanced glycation end products have been found to be associated with the formation of amyloid-ß and neurofibrillary tangles in AD. As a saccharification inhibitor, the glyoxalase system and its substrate methylglyoxal (MG) were certified to be associated with AD onset and development. As an active substance of AGEs, MG could cause direct or indirect damage to nerve cells and tissues. MG is converted to D-lactic acid after decomposition by the glyoxalase system. Under normal circumstances, MG metabolism is in a dynamic equilibrium, whereas MG accumulates in cells in the case of aging or pathological states. Studies have shown that increasing glyoxalase activity and reducing the MG level can inhibit the generation of oxidative stress and AGEs, thereby alleviating the symptoms and signs of AD to some extent. This paper focuses on the relevant mechanisms of action of the glyoxalase system and MG in the pathogenesis of AD, as well as the potential of inhibiting the production of advanced glycation end products in the treatment of AD.


Assuntos
Doença de Alzheimer/metabolismo , Córtex Cerebral/metabolismo , Lactoilglutationa Liase/metabolismo , Aldeído Pirúvico/metabolismo , Doença de Alzheimer/patologia , Animais , Córtex Cerebral/patologia , Produtos Finais de Glicação Avançada/metabolismo , Humanos , Estresse Oxidativo/fisiologia
12.
Zhonghua Nan Ke Xue ; 23(12): 1116-1120, 2017 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-29738185

RESUMO

Objective: To observe the effect of Huangjing Zanyu Capsule (HZC) on sperm mitochondrial membrane potential (MMP) in asthenozoospermia patients. METHODS: We assigned 70 asthenozoospermia patients to a treatment group (n = 39) and a control group (n = 31), the former treated with oral HZC at the dose of 4 capsules tid for 3 months while the latter left untreated. We obtained semen parameters from the patients and detected their sperm mitochondrial membrane potentials (MMP) by JC-1 staining and flow cytometry before and after medication, followed by comparison between the two groups. RESULTS: The total effectiveness rate was 71.05% in the treatment group and natural pregnancy was achieved in 3 cases during the medication. A total of 35 patients in the treatment group and 30 controls completed all the laboratory examinations after a 3-month observation. Compared with the controls, the patients treated with HZC exhibited significant improvement after medication in MMP (variation value: ï¼»1.19 ± 10.36ï¼½% vs ï¼»20.28 ± 14.21ï¼½%, P <0.01), total sperm motility (variation value: ï¼»3.46 ± 8.67ï¼½% vs ï¼»20.68 ± 14.12ï¼½%, P <0.01), the percentage of progressively motile sperm (variation value: ï¼»2.26 ± 8.29ï¼½% vs ï¼»17.58 ± 12.73ï¼½%, P <0.01), and the percentage of morphologically normal sperm (variation value: ï¼»0.23 ± 3.48ï¼½% vs ï¼»3.37 ± 3.99ï¼½%, P <0.01). MMP was significantly correlated with total sperm motility (r = 0.69, P <0.01), progressive sperm motility (r = 0.75, P <0.01) and normal sperm morphology (r = 0.26, P <0.01). CONCLUSIONS: Huangjing Zanyu Capsule can enhance sperm mitochondrial membrane potential and sperm mitochondrial function, thus improving total sperm motility, progressive sperm motility and normal sperm morphology. It is safe and effective for the treatment of asthenospermia.


Assuntos
Astenozoospermia/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Cápsulas , Estudos de Casos e Controles , Medicamentos de Ervas Chinesas/administração & dosagem , Feminino , Citometria de Fluxo , Humanos , Masculino , Potencial da Membrana Mitocondrial/fisiologia , Gravidez , Sêmen/efeitos dos fármacos , Motilidade Espermática/efeitos dos fármacos , Espermatozoides/ultraestrutura , Coloração e Rotulagem
13.
Tumour Biol ; 37(9): 12203-12211, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27230680

RESUMO

Prostaglandin E2 (PGE2), a derivative of arachidonic acid, has been identified as a tumorigenic factor in many cancers in recent studies. Prostaglandin E synthase 2 (PTGES2) is an enzyme that in humans is encoded by the PTGES2 gene located on chromosome 9, and it synthesizes PGE2 in human cells. In our study, we selected 119 samples from endometrial cancer patients, with 50 normal endometrium tissue samples as controls, in which we examined the expression of PTGES2. Both immunohistochemistry (IHC) and Western blot analyses demonstrated that synthase PTGES2, which is required for PGE2 synthesis, was highly expressed in endometrium cancer tissues compared with normal endometrium. Stable PTGES2-shRNA transfectants were generated in Ishikawa and Hec-1B endometrial cancer cell lines, and transfection efficiencies were confirmed by RT-PCR and Western blot analyses. We found that PGE2 promoted proliferation and invasion of cells in Ishikawa and Hec-1B cells by cell counting kit-8 tests (CCK8) and transwell assays, respectively. PGE2 stimulation enhanced the expression of SUMO-1, via PGE2 receptor subtype 4 (EP4). Further analysis implicated the Wnt/ß-catenin signaling pathway function as the major mediator of EP4 and SUMO-1. The increase in SUMO-1 activity prompted the SUMOlyation of target proteins which may be involved in proliferation and invasion. These findings suggest SUMO-1 and EP4 as two potential targets for new therapeutic or prevention strategies for endometrial cancers.


Assuntos
Proliferação de Células/efeitos dos fármacos , Dinoprostona/farmacologia , Neoplasias do Endométrio/metabolismo , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Proteína SUMO-1/metabolismo , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/genética , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Invasividade Neoplásica , Prostaglandina-E Sintases/genética , Prostaglandina-E Sintases/metabolismo , Interferência de RNA , Receptores de Prostaglandina E Subtipo EP4/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína SUMO-1/genética
14.
PLoS One ; 8(12): e81366, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24358111

RESUMO

Infection by human papillomavirus (HPV) can cause cervical intraepithelial neoplasia (CIN) and cancer. Down-regulation of E6 and E7 expression may be responsible for the positive clinical outcomes observed with IFN treatment, but the molecular basis has not been well determined. As miRNAs play an important role in HPV induced cervical carcinogenesis, we hypothesize that IFN-ß can regulate the expressions of specific miRNAs in cervical cancer cells, and that these miRNAs can mediate E6 and E7 expression, thus modulate their oncogenic potential. In this study, we found that miR-129-5p to be a candidate IFN-ß inducible miRNA. MiR-129-5p levels gradually decrease with the development of cervical intraepithelial lesions. Manipulation of miR-129-5p expression in Hela cells modulates HPV-18 E6 and E7 viral gene expression. Exogenous miR-129-5p inhibits cell proliferation in Hela cells, promotes apoptosis and blocks cell cycle progression in Hela cells. SP1 is a direct target of miR-129-5p in Hela cells. This study is the first report of a cellular miRNA with anti-HPV activity and provides new insights into regulatory mechanisms between the HPV and the IFN system in host cells at the miRNA level.


Assuntos
Proteínas de Ligação a DNA/genética , Regulação para Baixo , Regulação Viral da Expressão Gênica , Interferon beta/farmacologia , MicroRNAs/genética , Proteínas Oncogênicas Virais/genética , Fator de Transcrição Sp1/genética , Proliferação de Células , Neoplasia Intraepitelial Cervical/virologia , Feminino , Genes Virais , Células HeLa , Humanos , Neoplasias do Colo do Útero/virologia
15.
J Obstet Gynaecol Res ; 39(2): 549-54, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23006047

RESUMO

AIM: The BRCA1 promoter is hypermethylated in ovarian cancer patients. We postulated that this hypermethylation might be involved in ovarian cancer progression. METHODS: To confirm our hypothesis, tissue and serum samples were collected from ovarian carcinoma patients and categorized according to tumor stage. Healthy or benign ovarian disease tissue samples and corresponding serum samples were used as controls. Breast and ovarian cancer susceptibility gene 1 (BRCA1) promoter methylation levels were detected by real-time polymerase chain reaction (PCR). Real-time PCR was also used to evaluate BRCA1 gene expression, and Western blot was performed to assay the expression of BRCA1 protein. RESULTS: BRCA1 showed hypomethylation in 30 normal ovarian and 30 benign ovarian tumors, but showed hypermethylation or methylation in ovarian cancer patients. There was also a significant difference in the BRCA1 promoter methylation levels between different ovarian cancer stages. Compared to stage I and the control groups, there were higher BRCA1 promoter methylation frequencies in stage II and III ovarian cancers. BRCA1 methylation correlated with the loss of BRCA1 expression. BRCA1 promoter in stage I tumors showed hypomethylated. CONCLUSION: Promoter hypermethylation may act as a biomarker for sporadic ovarian cancer progression, but is unlikely to be helpful in the early diagnosis of ovarian cancer.


Assuntos
Proteína BRCA1/metabolismo , Metilação de DNA , Epigênese Genética , Neoplasias Ovarianas/metabolismo , Ovário/metabolismo , Regiões Promotoras Genéticas , Regulação para Cima , Proteína BRCA1/genética , Biomarcadores/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Estadiamento de Neoplasias , Neoplasias Ovarianas/patologia , Ovário/patologia
16.
Oncol Lett ; 3(4): 865-870, 2012 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-22741008

RESUMO

Special AT-rich sequence binding protein 1 (SATB1) is a master chromatin organizer that has recently been reported to directly upregulate metastasis-associated genes and downregulate tumor suppressor genes. However, its clinical significance in the case of ovarian cancer remains unclear. In the current study, we assessed the expression levels of SATB1 in ovarian cancer and aimed to show whether it may be a conventional clinicopathological parameter. Epithelial ovarian cancer (n=91), borderline cystadenoma (n=13) and normal ovarian background tissues (n=8) were collected immediately following excision during surgery. The mRNA expression levels of SATB1, VEGF-A and MMP-9 were determined using real-time quantitative PCR. Western blotting and immunohistochemical staining were carried out to detect the protein expression levels of SATB1. Expression levels within the ovarian cancer specimens were compared to the normal background tissues and analyzed against FIGO stage, lymph node involvement and histological type. SATB1 mRNA in malignant and borderline ovarian cystadenoma tissues was 6.74- and 5.70-fold higher compared with normal ovarian tissue, respectively (P<0.01). Western blot analysis revealed that a strong positive band of SATB1 expression was present in ovarian cancer tissues. Immunohistochemical staining showed that the positive expression rates of SATB1 in ovarian cancer, borderline ovarian cystadenoma and normal ovarian tissues were 69.2, 61.5 and 0% (P<0.01), respectively. SATB1 expression increased concomitantly with increasing FIGO stage and lymph node involvement. Survival curves showed that a higher SATB1 expression was correlated with shorter survival. Our results provide evidence that SATB1 expression is significantly associated with progression, metastasis and prognosis of epithelial ovarian cancer. SATB1 may therefore serve as a conventional clinicopathological parameter of ovarian cancer.

17.
J Cancer Res Clin Oncol ; 138(5): 775-83, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22270964

RESUMO

PURPOSE: The mechanisms underlying the effects of estrogen on endometrial cancer remain undefined. Although the classical mechanism of the action of estrogen involves binding to the estrogen receptors α and ß, and transduction of the signal into the cell, G protein-coupled receptor (GPR) 30 has been shown to mediate nongenomic estrogen signaling. The goal of this study was to determine the role of GPR30 signal in the basic process such as invasion and carcinogenesis of endometrial cancer. METHODS: We downregulated the expression of GPR30 in endometrial cancer cell line RL95-2 by transfection with shGPR30-pGFP-V-RS, a GPR30 antisense expression vector. The cells were then subjected to an MTT assay and a Transwell(®) migration assay. And an animal model was also used to investigate the influence of downregulation of GPR30 on oncogenesis. RESULTS: Downregulation of GPR30 led to reduced growth and invasion by cells treated with 17ß-estradiol. And the capacity of transfected RL95-2 cells to promote tumorigenesis was weakened in vivo. CONCLUSIONS: Our data suggest that, for the endometrial cancer cell line RL95-2, GPR30 plays important roles in mediating the proliferative and invasive effects of estrogen and in tumorigenesis.


Assuntos
Carcinoma/patologia , Transformação Celular Neoplásica/genética , Neoplasias do Endométrio/patologia , Receptores de Estrogênio/fisiologia , Receptores Acoplados a Proteínas G/fisiologia , Carcinoma/genética , Carcinoma/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Transformação Celular Neoplásica/efeitos dos fármacos , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/metabolismo , Estrogênios/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas de Membrana/fisiologia , Invasividade Neoplásica , RNA Interferente Pequeno/farmacologia , Receptores de Estrogênio/antagonistas & inibidores , Receptores de Estrogênio/genética , Receptores de Estrogênio/metabolismo , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Transfecção
18.
Cancer Lett ; 314(1): 41-53, 2012 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-21983130

RESUMO

This study investigated the role of miR-206 in estrogen receptor-α (ERα)-positive endometrial endometrioid adenocarcinoma (EEC). We profiled miR-206 expression in 30 EEC clinical samples using qRT-PCR, and explored its relationship with ERα and clinical parameters. A luciferase reporter assay assessed the ERα targeting potential of miR-206. Functional analyses of miR-206 were performed in EEC cell lines. MiRNA-206 expression decreased in ERα-positive EECs, and its expression was negatively correlated with ERα. MiRNA-206 overexpression inhibited ERα-dependent proliferation, impaired invasiveness and induced cell cycle arrest of ERα-positive EEC cell lines. Therefore, aberrantly expressed miRNA-206 may be associated with the development of ERα-positive EEC.


Assuntos
Carcinoma Endometrioide/patologia , Proliferação de Células , Neoplasias do Endométrio/patologia , Receptor alfa de Estrogênio/análise , MicroRNAs/fisiologia , Adulto , Carcinoma Endometrioide/química , Linhagem Celular Tumoral , Neoplasias do Endométrio/química , Receptor alfa de Estrogênio/genética , Receptor alfa de Estrogênio/fisiologia , Feminino , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica
19.
Int J Gynecol Cancer ; 21(8): 1357-65, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21720253

RESUMO

OBJECTIVE: The objectives of the study were to evaluate the role of mitogen-activated protein kinase (MAPK) signaling in normal, hyperplastic, and neoplastic endometrium in relation to estrogen receptor (ER) status and to investigate whether 17ß-estradiol (E2) and tamoxifen (TAM) mediate the proliferation and apoptosis of endometrial cancer cells through the MAPK pathway. METHODS: The expressions of phosphorylated and total extracellular signal-regulated kinases 1/2 (phosphorylated extracellular signal-regulated kinase 1/2 [p-ERK1/2] and total ERK1/2 [t-ERK1/2]) were analyzed with immunohistochemistry in normal, hyperplastic, and neoplastic endometrium. The expression levels of p-ERK1/2 and t-ERK1/2 in RL95-2 and KLE after stimulation by E2, progesterone (P), and TAM were detected by Western blotting. The effects of E2 and TAM in combination with MAPK pathway inhibitors on the growth and apoptosis of endometrial cancer cells were examined by the MTS assay and flow cytometry analysis. RESULTS: The expression level of p-ERK1/2 was significantly associated with the International Federation of Gynecology and Obstetrics stage (P = 0.0072). The ratio of phosphorylated/total ERK1/2 was higher in ER-positive endometrial cancer tissues and cells (P < 0.05). 17ß-Estradiol increased ERK1/2 phosphorylation, and TAM decreased ERK1/2 phosphorylation in endometrial cancer cell lines within 30 minutes (P < 0.05). The MEK1/2 inhibitor, U0126, and the stress-activated protein kinase/c-Jun NH2-terminal kinase inhibitor, SP600125, significantly suppressed the proliferation of human endometrial cancer cell lines RL95-2 and KLE induced by E2 (P < 0.05). The level of TAM-induced apoptosis was greater in KLE than in RL95-2 cells, and the p38 cascade was involved in the TAM-induced apoptosis of both cell lines (P < 0.05). CONCLUSIONS: The cross-talk between MAPK signaling and ER status might exert a key role in progression of endometrial cancer. Furthermore, the effects of E2 or TAM on the proliferation or apoptosis of ER-positive and ER-negative endometrial cancer cells were mediated through distinct MAPK pathways. These mechanisms might contribute to ER-specific differences in MAPK activation for molecular-target therapies in endometrial carcinoma.


Assuntos
Adenocarcinoma/etiologia , Neoplasias do Endométrio/etiologia , Sistema de Sinalização das MAP Quinases , Receptor Cross-Talk , Receptores de Estrogênio/metabolismo , Adenocarcinoma/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose , Estudos de Casos e Controles , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Neoplasias do Endométrio/metabolismo , Estradiol , Feminino , Humanos , Pessoa de Meia-Idade , Fosforilação , Tamoxifeno
20.
Int J Gynecol Cancer ; 21(2): 213-21, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21270604

RESUMO

INTRODUCTION: A long-term treatment with progestin commonly results in progestin resistance in endometrial cancer. So, the aim of this study was to investigate the role of glyoxalase I (GloI), a mediator of chemotherapy resistance, in metformin reversal of progestin resistance in endometrial carcinoma. METHODS: The proliferation variety of endometrial cancer cells was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium (MTT) assay after exposure to medroxyprogesterone acetate, metformin, or both reagents; apoptosis rates were assessed by flow cytometry. Real-time polymerase chain reaction was used to evaluate the effect of small interfering RNA sequence on target gene expression. Western immunoblotting was performed to determine the expression of GloI and the molecules of the mammalian target of rapamycin (mTOR) pathway. RESULT: Knocking down GloI sensitized progestin-resistant Ishikawa cells to progestin. Metformin downregulated GloI expression, reversed progestin resistance, enhanced progestin-induced cell proliferation inhibition, and induced apoptosis in progestin-resistant Ishikawa cells. In addition, medroxyprogesterone acetate-induced mTOR phosphorylation was blocked by metformin. Metformin abolishes mTOR phosphorylation and inhibits GloI expression, attenuating proliferation and inducing apoptosis in progestin-resistant Ishikawa cells. CONCLUSIONS: Dysregulation of GloI expression in endometrial cancer may be part of the molecular mechanisms for progestin resistance.


Assuntos
Carcinoma Endometrioide/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Neoplasias do Endométrio/tratamento farmacológico , Inibidores Enzimáticos/farmacologia , Lactoilglutationa Liase/metabolismo , Metformina/farmacologia , Progestinas/metabolismo , Apoptose , Carcinoma Endometrioide/metabolismo , Regulação para Baixo , Neoplasias do Endométrio/metabolismo , Feminino , Humanos , Células Tumorais Cultivadas
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