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1.
Gene ; 805: 145904, 2021 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-34418470

RESUMO

Breast cancer is the second most common cause of cancer-related mortality in women. Breast cancer metastasis which usually is observed at the last stage is the major cause of breast cancer-related death. Long non-coding RNAs (lncRNAs) are member of the non-coding RNA family. It is known that lncRNAs have important functions in the genes regulation of different processes and pathways such as EMT (Epithelial mesenchymal transition), metastasis and apoptosis. Therefore, it is inevitable that lncRNAs have potential contribution for the understanding of cancer pathogenesis. lncRNA-ZEB2NAT is the natural antisense transcript of ZEB2. Herein, we investigated the effects of lncRNA-ZEB2NAT on process of EMT, metastasis and apoptosis in MCF7 and MDA-MB-231 breast cancer cells. The effect of ZEB2NAT on the expression of important genes in EMT, metastasis and apoptosis, and some protein levels was determined by qRT-PCR and western blot analysis, respectively. The effects of ZEB2NAT on cell proliferation, apoptosis, invasion and colony formation were evaluated using XTT, annexin V, invasion and colony assays, respectively. The ZEB2NAT knockdown caused anti-metastatic and apoptotic effects. The ZEB2NAT knockdown resulted in a decrease in ZEB2 and N-cadherin but an increase in E-cadherin protein levels. In addition, it was determined that ZEB2NAT knockdown significantly decreased cell proliferation and stimulated apoptosis in both cells. It was found that ZEB2NAT knockdown significantly decreased invasion and colony formation in both cells. ZEB2NAT knockdown showed anti-metastatic and apoptotic effect by affecting the important genes in both cells. These results have suggested that ZEB2NAT has an important role in EMT, metastasis and apoptosis in breast cancer and ZEB2NAT knockdown caused significant anti-cancer activities.


Assuntos
Neoplasias da Mama/genética , Homeobox 2 de Ligação a E-box com Dedos de Zinco/genética , Apoptose/genética , Neoplasias da Mama/metabolismo , Caderinas/genética , Caderinas/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Transição Epitelial-Mesenquimal/genética , Feminino , Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Invasividade Neoplásica/genética , Metástase Neoplásica/genética , RNA Longo não Codificante/genética , Homeobox 2 de Ligação a E-box com Dedos de Zinco/metabolismo
2.
Med Oncol ; 38(9): 113, 2021 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-34378101

RESUMO

The aim of this study was to determine the effect of lncRNA HIF1A-AS2 on autophagy-associated drug resistance in small cell lung cancer (SCLC) cells. The expression of HIF1A-AS2 was silenced by siRNA in doxorubicin-sensitive H69 and doxorubicin-resistant H69AR cells. Then, cytotoxicity, apoptosis and autophagy analyses were carried out in the normoxic and CoCl2-induced hypoxic environment. The effect of HIF1A-AS2 on the expression levels of genes, which are associated with drug resistance and autophagy, was determinated by qRT-PCR analysis. The levels of MRP1, HIF-1α and Beclin-1 were analyzed by western blot method. Knockdown of HIF1A-AS2 increased doxorubicin sensitivity of SCLC cells and decreased autophagy. Knockdown of HIF1A-AS2 has also affected the expression of several genes that will increase drug sensitivity and inhibit autophagy in both cell lines. The levels of HIF-1α and Beclin-1 were decreased in both cell lines by knockdown of HIF1A-AS2. MRP1 expression was decrease in H69AR cells. In addition, CoCl2-induced hypoxic environment decreased in doxorubicin sensitivity of H69 cells, and knockdown of HIF1A-AS2 reversed this effect of hypoxia. Knockdown of HIF1A-AS2 increased drug sensitivity of SCLC cells in relation to autophagy. Therefore, hypoxia-HIF1A-AS2-autophagy interaction is thought to be determinative in drug sensitivity of these cells.

3.
Int J Psychiatry Clin Pract ; 24(2): 120-126, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31916884

RESUMO

Objective: Suicide is the second leading cause of death in adolescence. Genetic studies implicate the genetic component of suicide independent from associated psychiatric disorder. Although genetics is an important factor that might be associated with suicide, limited progress is achieved to identify the candidate genes in adolescents.Methods: The study included 97 patients and 106 controls. Five leptin receptor single nucleotide polymorphism (SNP) region examined. Depressive scores were measured with Children Depression Inventory and suicidal behaviour was measured by Suicide Probability Scale. Logistic and linear regression analysis used for determining to predictors.Results: In linear regression analysis (R2: 0.786) both previous suicide attempt (B:5.553, t:2.613 p: .035) and having a mutant allele in rs1171276 SNP region (B:4.346 t:2.220 p: .048) have been associated with suicidal behaviour. In logistic regression analysis, family history of depression (p < .0001, OR: 4.2 [1.7-9.6]) and number of stressful life events (p: .001, OR: 1.7 [1.3-2.1]) predicted depression significantly.Conclusion: Leptin receptor polymorphism could result in an increase in impulsive behaviour and suicide scores with leptin resistance. Our research is the first study to investigate the relationship between depression, suicidal behaviour and leptin receptor polymorphism in adolescent sample. Similar studies could be carried out on a community basis.KEY POINTSAn association has been found between rs1171276 single nucleotide polymorphism of leptin receptor and the suicide probability scores in depressed adolescents.Family history of depression and number of stressful life events predict depressive scores significantly.Leptin receptor single nucleotide polymorphism may be a risk factor for adolescent suicide by increasing impulsive behaviour.


Assuntos
Comportamento do Adolescente/fisiologia , Transtorno Depressivo/fisiopatologia , Comportamento Impulsivo/fisiologia , Receptores para Leptina/genética , Estresse Psicológico/fisiopatologia , Tentativa de Suicídio , Adolescente , Transtorno Depressivo/genética , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Polimorfismo de Nucleotídeo Único , Fatores de Risco
5.
Gene ; 671: 127-134, 2018 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-29792952

RESUMO

Sinapic acid (SA) is a derivative of hydroxycinnamic acid and found in various vegetables and fruit species. Aim was to evaluate the anticancer effects of SA in PC-3 and LNCaP human prostate cancer cells. The effect of SA on cell viability was determined using XTT assay. Expressions of 8 genes for apoptosis and 6 genes for metastasis were evaluated by qPCR. Caspase-3 activity was determined using caspase-3 colorimetric assay kit. Effect of SA on cell invasion was evaluated with cell invasion assay. The IC50 dose of SA in PC-3 and LNCaP cells was found to be 1000 µM for 72 h. SA treatment increased the expression of BAX, CASP3, CASP8, CYCS, FAS, TIMP-1 and CDH1 however significantly decreased the expression of MMP-9 in PC-3 cells. In LNCaP cells, the expressions of BAX, CASP3, CASP7 and CYCS were significantly elevated; however, a decrease was seen in the expressions of CDH2, MMP-2 and MMP-9 in the SA treatment. Moreover, SA significantly increased caspase-3 activity and suppressed the cell invasion. In conclusion, it is thought that SA has anticancer effect on prostate cancer cells. However, more detailed studies should be conduct to illuminate molecular mechanism of apoptotic and antimetastatic activity of SA.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Biomarcadores Tumorais/genética , Ácidos Cumáricos/farmacologia , Neoplasias da Próstata/genética , Proteínas Reguladoras de Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Masculino , Neoplasias da Próstata/tratamento farmacológico
6.
Biomed Res Int ; 2016: 2830394, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27092309

RESUMO

The objective of this study was to determine the intra- and intergenetic diversities of eight different goat populations in Turkey including Hair, Angora, Kilis, Yayladag, Shami, Honamli, Saanen, and Alpine. A total of 244 DNA samples were genotyped using 11 microsatellites loci. The genetic differentiation between breeds was considerable as a result of the statistically significant (P < 0.001) pairwise F ST values of each pair of breeds. Exceptionally, F ST values calculated for Honamli and Hair breeds were statistically nonsignificant (P > 0.05). Heterozygosity values ranged between 0.62 and 0.73. According to the structure and assignment test, Angora and Yayladag goats were assigned to the breed they belong to, while other breeds were assigned to two or more different groups. Because this study for the first time presented genetic data on the Yayladag goat, results of structure analysis and assigned test suggest that further analyses are needed using additional and different molecular markers.


Assuntos
Variação Genética , Cabras/genética , Repetições de Microssatélites/genética , Animais , Cruzamento , Genótipo , Turquia
7.
Theriogenology ; 85(4): 638-44, 2016 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-26559469

RESUMO

Early pregnancy is one of the most critical periods of pregnancy, and many factors such as cytokines, enzymes, and members of the immune system have to cooperate in a balanced way. In the present study, the gene expression profiles of factors associated with pregnancy such as EGF, transforming growth factor beta, granulocyte-macrophage colony-stimulating factor, interferon gamma, insulin-like growth factor 2, insulin-like growth factor 2 receptor, and matrix metalloproteinase 2 were analyzed in uterine tissues of female cats. The cats were assigned to five groups: G1 (embryo positive, n = 7; 7th day after mating), G2 (after implantation, n = 7; 20th day after mating), G3 (midgestation, n = 7; 24-25th day after mating), G4 (late gestation, n = 7; 30-45th day after mating), G5 (oocyte group, n = 7; 7th day after estrus). Tissue samples from the uterus and placenta were collected after ovariohysterectomy. Relative messenger RNA levels were determined by real-time polymerase chain reaction. All the factors examined were detected in all tissue samples. In the course of pregnancy, significantly higher expression of EGF and matrix metalloproteinase 2 in G2 than in G1 was observed (P < 0.05). Insulin-like growth factor 2 expression was higher in all groups than in G1 (P < 0.05). Upregulation of EGF during implantation was detected. The expression of interferon gamma was significantly higher in G3 than in G1 (P < 0.05). Transforming growth factor beta and granulocyte-macrophage colony-stimulating factor were constantly expressed in all groups. In conclusion, the expressions of these factors in feline uterine tissue at different stages of pregnancy might indicate that these factors play roles in the development of pregnancy such as trophoblast invasion, vascularization, implantation, and placentation.


Assuntos
Gatos/fisiologia , Citocinas/metabolismo , Enzimas/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Prenhez , Transcriptoma , Animais , Citocinas/genética , Enzimas/genética , Feminino , Regulação da Expressão Gênica/fisiologia , Peptídeos e Proteínas de Sinalização Intercelular/genética , Gravidez , Prenhez/fisiologia
8.
Theriogenology ; 84(1): 24-33, 2015 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-25794839

RESUMO

Hypoxia-inducible factors (HIFs) and vascular endothelial growth factor (VEGF) have critical roles during the development of the fetomaternal unit. The HIFs regulate placentation and vascularization by stimulation of VEGF gene expression. This study aimed to investigate the expression profiles of HIF gene family and VEGF in the cat uterus during pregnancy. Tissue samples of the whole uterine wall were collected after ovariohysterectomy and allocated to the following groups: embryo positive (group 1 [G1], n = 7, 7 days after mating), early pregnancy (group 2 [G2], n = 7, 20 days after mating), mid-pregnancy (group 3 [G3], n = 7, 24 days after mating), late pregnancy (group 4 [G4], n = 7, 30-45 days after mating), and oocyte positive groups (group 5 [G5], n = 7, 7 days after induction of ovulation with GnRH analog). Relative mRNA levels were determined by real-time polymerase chain reaction. As housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase was used. The relative gene expression of HIF1A in G5 was found to be significantly higher than that of other groups (G1, G2, G3, and G4) (P < 0.05). In addition, the expression of HIF2A in G5 was higher than that of G1 and HIF2A gene expression at placentation sites of G4 was higher than in G1, G2, and G3 (P < 0.05). Immunohistochemistry indicated that HIF1A, HIF2A, and VEGF expressions were observed in different cell types of uterine and placental tissues in late pregnancy and oocyte groups. The expression of HIF3A did not change significantly in any group investigated. These observations suggest that HIFs and VEGF may play a role in the establishment and development of pregnancy.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Gatos/fisiologia , Prenhez/metabolismo , Útero/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/análise , Gatos/metabolismo , Feminino , Imuno-Histoquímica , Gravidez , RNA Mensageiro/metabolismo , Fator A de Crescimento do Endotélio Vascular/análise
9.
Angle Orthod ; 84(5): 803-9, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24654940

RESUMO

OBJECTIVE: To evaluate the allele and genotype frequencies of the P561T and C422F polymorphic sites of the growth hormone receptor (GHR) gene and the relationship between mandibular prognathism (MP) and these two single-nucleotide polymorphisms (SNPs). MATERIALS AND METHODS: A total of 99 subjects with severe skeletal Class III MP who planned to undergo orthognathic surgery and 99 subjects with Class I occlusion were examined in this study to evaluate the relationship between MP and two SNPs in exon 10 of the GHR gene. GHR was chosen as a candidate gene because growth hormone plays an important role in cartilage growth. A blood sample was used to extract genomic DNA, and the polymerase chain reaction-restriction fragment length polymorphism method was used to determine genotypes of P561T and C422F. The Minitab 14.0 packet program was used to perform statistical analysis. RESULTS: Allele frequencies of the C422F and P561T variants were determined. Because of the low allele frequency of the control group, statistical analysis could not be performed to test the difference between MP and control groups. Therefore, the data were combined to determine the association between the P561T polymorphism and craniofacial measurements. Effective mandibular length (condylion-gnathion) and lower face height (anterior nasal spina-menton) were associated with the P561T variant. CONCLUSION: This finding supports that the GHR might be a candidate gene for mandibular morphogenesis in this population.


Assuntos
Cisteína/genética , Fenilalanina/genética , Polimorfismo de Nucleotídeo Único/genética , Prognatismo/genética , Prolina/genética , Receptores da Somatotropina/genética , Treonina/genética , Cefalometria/métodos , Queixo/patologia , Códon/genética , Éxons/genética , Feminino , Frequência do Gene/genética , Genótipo , Haplótipos/genética , Heterozigoto , Humanos , Masculino , Má Oclusão Classe III de Angle/genética , Mandíbula/patologia , Côndilo Mandibular/patologia , Maxila/patologia , Osso Nasal/patologia , Polimorfismo de Fragmento de Restrição/genética , Dimensão Vertical , Adulto Jovem
10.
Theriogenology ; 80(2): 145-52, 2013 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-23623165

RESUMO

The aims of the present study were to elucidate the expression profiles of leukotriene (LT) pathway mRNA transcription and to determine the possible interaction of LT and prostaglandin (PTG) pathways genes in equine endometrium during the estrous cycle and early pregnancy. Endometrial biopsies were obtained from mares on the day of ovulation (d0), at late diestrous (LD, n = 4), and after luteolysis in the estrus phase (AL, n = 4) of the cycle. Biopsies were also taken on Days 14 (P14; n = 4), 18 (P18, n = 4), and 22 (P22, n = 4) during early pregnancy that were comparable days to cyclic sampling days. A mixed model was fitted on the normalized relative mRNA levels, quantified by qPCR in duplicate, and least significant difference test was employed to detect significantly different group(s). In addition, to determine the degree of contribution of each gene to separation of treatment groups, the multivariate projection method partial least square regression discriminant analysis was used. The expression of 5-lipoxygenase mRNA was greater on d0 and LD, declined at AL, and was suppressed by early pregnancy. Leukotriene A4 hydrolase mRNA expression increased at LD and during early pregnancy, but was significantly greater at LD compared with P14. The expression of LT C4 synthase mRNA was only induced at LD. Cysteinyl leukotriene receptors (CysLT1 and CysLT2) mRNA expressions were decreased by both cyclic changes and early pregnancy, whereas 5-lipoxygenase-activating protein and B leukotriene receptor mRNA expressions were not affected by early pregnancy or stages of the estrous cycle. Partial least square discriminant analysis suggests that LT and PTG pathway enzymes and receptors appear to behave similarly in terms of mRNA expression. In conclusion, the expression profiles of LT pathway genes are demonstrated in equine endometrium for the first time by the present study, and the present data suggest that LT pathway mRNA transcriptions are tightly regulated during early pregnancy in mares.


Assuntos
Endométrio/metabolismo , Enzimas/genética , Ciclo Estral/genética , Cavalos/fisiologia , Leucotrienos/metabolismo , Prenhez , Receptores de Leucotrienos/genética , Animais , Ácido Araquidônico/metabolismo , Enzimas/metabolismo , Ciclo Estral/sangue , Feminino , Idade Gestacional , Masculino , Gravidez , Prenhez/genética , Receptores de Leucotrienos/metabolismo , Transdução de Sinais/genética
11.
Anim Reprod Sci ; 125(1-4): 124-32, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21411251

RESUMO

The aim was an evaluation of a set of housekeeping genes (HKGs) to be used in the normalization of gene expression in the equine endometrium. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine ribosyl transferase 1 (HPRT1), ubiquitin B (UBB), tubulin alpha 1 (TUBA1), ribosomal protein L32 (RPL32), beta-2-microglobulin (B2M), 18S rRNA (18S), and 28S rRNA (28S) HKGs were evaluated using real-time PCR and were compared in different physiological stages of the endometrium. Endometrial biopsies were obtained from mares on day of ovulation (d0, n=4), at late diestrus (LD, n=4), after luteolyis (AL, n=4) of the cycle and on days 14 (P14; n=3), 18 (P18, n=3) and 22 (P22; n=3) of pregnancy. A model based on REML with support of descriptive statistics was proposed in accordance with experimental design and was further confirmed with principal component analysis (PCA). Results were compared with widely used software including geNorm, BestKeeper, and NormFinder. Results indicated that GAPDH was the most stable HKG and RPL32 was ranked as the second best. 18S and 28S were found to be the least stable. The proposed model, PCA, geNorm, and BestKeeper were in agreement in detecting the most stable and the least stable HKGs in the equine endometrium during the estrous cycle and early pregnancy.


Assuntos
Endométrio/fisiologia , Ciclo Estral/genética , Cavalos/genética , Prenhez/genética , Animais , Endométrio/metabolismo , Feminino , Perfilação da Expressão Gênica/métodos , Perfilação da Expressão Gênica/veterinária , Modelos Lineares , Gravidez , Análise de Componente Principal , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária
12.
Anim Reprod Sci ; 122(1-2): 124-32, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20832957

RESUMO

The aim was to evaluate expression of genes involved in the biosynthesis of prostaglandins (PTG), Prostaglandin H Synthase-1 (PTGS1) and PTGS2, PGF synthase (PTGFS), and PGE synthase (PTGES), PGF receptor (PTGFR), PGE receptors (PTGER2 and PTGER4), prostaglandin transporter (SLCO2A1) and hydroxyprostaglandin dehydrogenase-15 (HPGD). Endometrial biopsies were obtained from mares on day of ovulation (d0, n=4), late diestrus (LD, n=4), early luteolysis (EL, n=4) and after luteolysis (AL, n=4) during the cycle. Stages of the cycle were confirmed by plasma progesterone concentrations measured daily and ultrasound examinations. Biopsies were also taken on days 14 (P14; n=4), 15 (P15, n=4), 18 (P18, n=4) and 22 (P22; n=4) of pregnancy. Relative mRNA expressions were quantified using real-time RT-PCR. A mixed model was fitted on the normalized data and least significant difference test (α=0.05) was employed. Expression of PTGS1 mRNA was low throughout the estrous cycle and early days of pregnancy, but upregulated on P18 and P22. PTGS2 expression was increased on EL, but it was suppressed by pregnancy on P15, P18, and P22. PTGFS expression was upregulated in both cyclic and pregnant mares compared to d0 and its level was the highest on LD. PTGFR expression was transiently increased on LD and EL and was suppressed during early pregnancy. Both PTGES and PTGER2 expressions were increased on LD, EL, and early pregnancy, but were decreased after the luteolysis in cyclic mares as they remained high on P18 and P22. PTGER4 expression did not change throughout the cycle and early pregnancy. Levels of HPGD and SLCO2A1 were significantly increased only on P22. In conclusion, PTGS2 expression increases around the time of luteolysis and concurrent upregulation of PTGFS and PTGES indicates that equine endometrium has increased capability of PTG production around the time of luteolysis. However, pregnancy reduces PTGS2 expression, but maintains the high levels of PTGES during early pregnancy along with PTGER2 while PTGFR expression was suppressed. These findings suggest that possible luteotrophic action of PGE2 is required in early equine pregnancy. PTGS1 is only upregulated later in the early pregnancy suggesting that it is not involved in luteolysis, but could be the main PTGS enzyme at this time during early pregnancy. An increase in HPGD and SLCO2A1 levels on P22 indicates a tight regulation of PTG action by pregnancy.


Assuntos
Endométrio/metabolismo , Ciclo Estral/genética , Regulação da Expressão Gênica no Desenvolvimento , Cavalos/metabolismo , Prenhez/genética , Prostaglandinas/genética , Animais , Endométrio/diagnóstico por imagem , Feminino , Hidroxiprostaglandina Desidrogenases/genética , Luteólise/genética , Luteólise/metabolismo , Transportadores de Ânions Orgânicos/genética , Gravidez , Progesterona/sangue , Prostaglandina-Endoperóxido Sintases/genética , Receptores de Prostaglandina/genética , Ultrassonografia
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