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Objective: To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer. Methods: A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot. Results: The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue (P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue (P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage (P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression (P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells (P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells (P=0.006 and P=0.030, respectively). Conclusions: HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.
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Carcinoma , Fator 4 Nuclear de Hepatócito/fisiologia , Neoplasias Gástricas , Linhagem Celular Tumoral , Proliferação de Células , Gastrectomia , Regulação Neoplásica da Expressão Gênica , Fator 4 Nuclear de Hepatócito/genética , Fator 4 Nuclear de Hepatócito/metabolismo , Fatores Nucleares de Hepatócito , Humanos , Prognóstico , Neoplasias Gástricas/genética , Neoplasias Gástricas/cirurgiaRESUMO
Objective: To explore the roll and function of hydroxymethylglutaryl-CoA synthase 2 (HMGCS2) in the development and progression of human esophageal squamous cell carcimoma(ESCC). Method: Using immunohistochemistry, the expression of HMGCS2 was determined in 150 primary ESCC patients from July 2002 to December 2005 in the People's Hospital of Linzhou City, Henan Province. And HMGCS2 over-expression ESCC cell lines were established to verify HMGCS2 gene function. Result: In 150 cases of ESCCs, the expression rate of HMGCS2 was 58% (87/150), which was lower than 72% (108/150) in paired normal tissues, the difference was statistically significant (P=0.013). HMGCS2 down-regulated expression was associated with tumor cell differentiation (P=0.022), pT status (P=0.036), pN status (P=0.017) and TNM stage(P=0.012). The 5-years disease-specific survival (DSS) in down HMGCS2 expression group (14 months) was poorer than those in normal expression group (20 months; P=0.002). In addition, multivariate Cox regression analysis showed that HMGCS2 expression (Wald=7.136, P=0.008) was an independent risk factor for DSS. Furthermore, functional studies demonstrated that HMGCS2 gene could suppress the tumorigenic ability of ESCC cells (OD: 0.79±0.04 vs 1.25±0.68; P=0.01), the formation of colone (number of colones: 30±10 vs 189±15, P=0.002), and cell motility (number of cells: 27±14 vs 222±40, P=0.009). Conclusion: HMGCS2 can inhibit the proliferation and migration of ESCC cells, and could be an important candidate tumor suppressor gene for ESCC.
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Neoplasias Esofágicas , Carcinoma de Células Escamosas do Esôfago , Biomarcadores Tumorais , Carcinoma de Células Escamosas , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Humanos , Hidroximetilglutaril-CoA Sintase , PrognósticoRESUMO
Immunological microenvironment is not only composed of multiple immune cells, but also deposited various inflammation factors that regulate immune response to tumor cells. To ascertain the crucial immune factors presented in hepatocellular carcinoma microenvironment (HCM), tumor tissue culture supernatant (TCS) and the corresponding non-tumor tissue culture supernatant (NCS) from patient with hepatocellular carcinoma (HCC) were analyzed by antibody array technology. Among the inflammation-associated cytokines assayed, high level of chemokines CXCL8/IL-8 (6.82-fold increase) and CXCL10/IP-10 (16.45-fold increase) in TCS than that in paired NCS were evidently identified. And low expression of IL-16 (0.14-fold decrease) and RANTES/CCL5 (0.17-fold decrease) in TCS were also uncovered. Especially, overexpression of CXCL10 in primary HCC compared with their non-tumor counterparts was significantly associated with serum AFP level (P = 0.004), tumor size (P = 0.021), tumor number (P < 0.001) and TNM stage (P = 0.027). In addition, Kaplan-Meier curves demonstrated that patients with higher CXCL10 expression levels had significantly poorer overall survival (P = 0.016) and disease-free survival (P = 0.022) than those with lower CXCL10 expression levels. Univariate and multivariate analyses revealed that the level of CXCL10 expression was an independent prognostic factor for overall survival in HCC patients. In summary, high concentration of CXCL10 is deposited in HCM identified by antibody array, which may contribute to the prediction of clinical outcome of HCC patients.
Assuntos
Carcinoma Hepatocelular/genética , Quimiocina CXCL10/genética , Neoplasias Hepáticas/genética , Microambiente Tumoral , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/diagnóstico , Humanos , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/diagnóstico , PrognósticoRESUMO
It has been confirmed that trimethylation of lysine 27 on histone H3 (H3K27me3) plays an important role in epigenetic process of tumorigenesis. However, the status of H3K27me3 in ovarian cancer and its impact on patients' clinicopathologic characteristics and prognosis are unclear. In the present study, the immunohistochemistry (IHC) was utilized to detect protein expression of H3K27me3 in 12 normal ovaries, 26 ovarian cystadenomas, 31 borderline ovarian tumors and 168 ovarian carcinomas by tissue microarray. The association between H3K27me3 expression with clinicopathologic features and patient prognosis were also evaluated using various statistical models. The expression of H3K27me3 was decreased in 2 of 12 (16.7%) cases of the normal ovaries, 8 of 26 (30.8%) cases of cystadenomas, 12 of 31 (38.7%) cases of borderline ovarian tumors, and 93 of 168 (55.4%) cases of primary ovarian carcinomas, respectively (P<0.05). Further correlation analysis suggested that decreased expression of H3K27me3 in ovarian carcinomas was significantly correlated with more advanced pM and FIGO stages (P<0.05). In addition, a significant association between decreased expression of H3K27me3 and shortened patient survival (mean 66 months versus 101 months, p=0.019) was demonstrated by univariate survival analysis of the ovarian carcinoma cohorts. Importantly, H3K27me3 expression provided a significant independent prognostic factor in multivariate analysis (p=0.028). These findings confirmed that decreased expression of H3K27me3 in primary ovarian cancer might be correlated with the acquisition of an invasive and/or aggressive phenotype of tumor, and might serve as an independent biomarker for poor prognosis in patients with ovarian carcinoma.
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AIM: To investigate the role of p300 in the regulation of proliferation and odontogenic differentiation of human dental pulp cells (HDPCs). METHODOLOGY: The recombinant lentiviral vector pshRNA-copGFP was used to knock-down p300 expression in HDPCs. Protein level of acetylated H3 was detected. The proliferation of HDPCs was measured using the CCK8 assay. The cell cycle and apoptosis were analysed using flow cytometry and TUNEL staining, respectively. The expression levels of Cdc25A, p21(waf1) and the cleaved products of caspase 3 and caspase 7 were determined utilizing real-time quantitative polymerase chain reaction and Western blotting analysis. The alkaline phosphatase (ALP) activity was measured, and the formation of mineralized nodules was assessed using alizarin red staining after the induction of odontogenic differentiation of HDPCs. The expression levels of the odontogenic differentiation markers DMP-1, DSPP and DSP were detected utilizing real-time quantitative polymerase chain reaction and Western blotting analysis. RESULTS: After p300 was knocked down in HDPCs, p300 was significantly down-regulated at both the mRNA and protein levels, and histone H3 acetylation was reduced. The proliferation capacity of HDPCs was suppressed in p300 knock-down groups. The cells were arrested in the G0/G1 phase of the cell cycle, and cell apoptosis was triggered. ALP activity, the formation of mineralized nodules and the expression levels of DMP-1, DSPP and DSP were all decreased in p300-knock-down HDPCs undergoing odontogenic differentiation. CONCLUSION: Knocking down p300 restrains the proliferation and odontogenic differentiation potentiality of HDPCs.
Assuntos
Ciclo Celular/fisiologia , Polpa Dentária/citologia , Proteína p300 Associada a E1A/fisiologia , Odontogênese/fisiologia , Adolescente , Adulto , Fosfatase Alcalina/metabolismo , Apoptose , Biomarcadores/metabolismo , Western Blotting , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Proteína p300 Associada a E1A/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Citometria de Fluxo , Humanos , Marcação In Situ das Extremidades Cortadas , Técnicas In Vitro , Fosfoproteínas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Sialoglicoproteínas/metabolismoRESUMO
Terminal deletions are found frequently in both malignancies and clinically recognizable deletion syndromes in man. Little is known, particularly in cancer, of the specific mechanisms which lead to the generation of deleted chromosomes or the process by which these broken chromosomes are stabilized. We demonstrate that several examples of apparent terminal deletions are, in fact, subtelomeric translocations which were not detectable using conventional cytogenetics. The unexpectedly high frequency of this phenomenon and the diversity of partner chromosomes involved in the subtelomeric translocations is consistent with a model in which telomere capture can stabilize chromosome breakage in man.
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Deleção Cromossômica , Telômero/ultraestrutura , Cromossomos Humanos Par 6 , Humanos , Células Híbridas/ultraestrutura , Hibridização in Situ Fluorescente , Melanoma/genética , Melanoma/ultraestrutura , Modelos Genéticos , Translocação GenéticaRESUMO
The strategy presented here to identify unequivocally cryptic chromosomal rearrangements has relevance to both prenatal and postnatal cytogenetic analysis as well as the analysis of tumour-associated chromosome rearrangements. Microdissection and in vitro amplification of specific chromosomal regions are performed, followed by labelling for fluorescent in situ hybridization (FISH) to normal metaphase chromosomes (Micro-FISH). Micro-FISH probes have been used successfully to determine the derivation of chromosome segments unidentifiable by standard chromosome banding analysis. Micro-FISH probes (created in less than 24 hours) now make it possible to identify explicitly the chromosome constitution of virtually all cytologically visible chromosome rearrangements.
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Cromossomos Humanos/ultraestrutura , Hibridização in Situ Fluorescente/métodos , Técnicas de Sonda Molecular , Sequência de Bases , Aberrações Cromossômicas , Deleção Cromossômica , DNA/genética , Sondas de DNA , Feminino , Rearranjo Gênico , Humanos , Dados de Sequência Molecular , Neoplasias/genética , Reação em Cadeia da Polimerase , Gravidez , Diagnóstico Pré-Natal , Translocação GenéticaRESUMO
We have performed microdissection of 16 putative homogeneously staining regions (hsrs) from nine different breast cancer cell lines in order to determine their chromosomal origin and composition. As expected, the most commonly amplified chromosomal band-region was 17q12 (containing ERBB2). However, regions not containing known oncogenes were also identified, including 13q31 (5/9 cases) and 20q12-13.2 (4/9 cases). The chromosomal composition of the integrated amplified DNA within each hsr was determined and in 13/16 cases (81%), hsrs were shown to be composed of two or more chromosomal regions. These studies shed light on the mechanism of formation of hsrs, and identify chromosomal regions likely to harbour genes amplified in breast cancer.
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Neoplasias da Mama/genética , Cromossomos Humanos/ultraestrutura , Amplificação de Genes , Hibridização in Situ Fluorescente/métodos , Micromanipulação , Sequência de Bases , Bandeamento Cromossômico , Cromossomos Humanos Par 13/ultraestrutura , Cromossomos Humanos Par 17/ultraestrutura , Cromossomos Humanos Par 20/ultraestrutura , Feminino , Humanos , Dados de Sequência Molecular , Oncogenes , Células Tumorais CultivadasRESUMO
Matrix metallopeptidase 10 (MMP10) is frequently expressed and correlates closely with metastasis and poor prognosis in various human cancers. However, the significance of MMP10 expression in esophageal squamous cell carcinoma (ESCC) and its role in ESCC progression remains unclear. In this report, upregulation of MMP10 mRNA was detected in 39/60 (65.0%) of primary ESCC tissues compared with their paired nontumor esophageal tissues. Tissue microarray (TMA) study found protein overexpression of MMP10 in 188/239 (78.7%) of primary ESCC tissues but not in their corresponding nontumor esophageal tissues, suggesting that overexpression of MMP10 may play important roles in ESCC development and progression. Although the overexpression of MMP10 was not significantly associated with disease-specific survival rate (P= 0.182) for all tested ESCCs, it was significantly associated with poorer disease-specific survival (P= 0.001) in early stage of ESCCs (I-IIA). In addition, multivariate analysis found that MMP10 expression in tumor tissues was evaluated as a potential independent prognostic factor for early stage ESCC patients. These findings suggest that MMP10 plays an important role in ESCC progression in the early stage, and overexpression of MMP10 in tumor tissues could be used as a potential prognostic marker for patients with early clinical stage of ESCC.
Assuntos
Carcinoma de Células Escamosas/diagnóstico , Neoplasias Esofágicas/diagnóstico , Regulação Neoplásica da Expressão Gênica , Metaloproteinase 10 da Matriz/metabolismo , RNA Mensageiro/análise , Regulação para Cima , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/metabolismo , Carcinoma de Células Escamosas do Esôfago , Feminino , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Masculino , Metaloproteinase 10 da Matriz/genética , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise Serial de TecidosRESUMO
Background and Purpose: Functional imaging has an established role in therapeutic monitoring of cancer treatments. This study evaluated the correlations of tumour permeability parameters derived from dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and tumour cellularity derived from apparent diffusion coefficient (ADC) in nasopharyngeal carcinoma (NPC). Material and Methods: Twenty NPC patients were examined with DCE-MRI and RESOLVE diffusion-weighted MRI (DW-MRI). Tumour permeability parameters were quantitatively measured with Tofts compartment model. Volume transfer constant (Ktrans), volume of extravascular extracellular space (EES) per unit volume of tissue (Ve), and the flux rate constant between EES and plasma (Kep) from DCE-MRI scan were measured. The time-intensity curve was plotted from the 60 dynamic phases of DCE-MRI. The initial area under the curve for the first 60 s of the contrast agent arrival (iAUC60) was also calculated. They were compared with the ADC value derived from DW-MRI with Pearson correlation analyses. Results: Among the DCE-MRI permeability parameters, Kep had higher linearity in inverse correlation with ADC value (r = -0.69, p = <0.05). Ktrans (r = -0.60, p=<0.05) and iAUC60 (r = -0.64, p = <0.05) also had significant inverse correlations with ADC. Ve showed a significant positive correlation with ADC (r = 0.63, p = <0.05). Conclusions: Nasopharyngeal tumour vascular permeability parameters derived from DCE-MRI scan were correlated linearly with tumour cellularity measured by free water diffusability with ADC. The clinical implementations of these linear correlations in the quantitative assessments of therapeutic response for NPC patients may be worth to further explore.
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Objective: To investigate the prevalence of five specific periodontal pathogens in the saliva of edentulous patients and to compare the differences in the saliva of dentulous individuals with various periodontal conditions. Methods: All the subjects were patients who received regular care at the Beijing Hypertension Prevention and Management Institute. Twenty-seven edentulous patients (edentulous group) were included. According to age (age gap≤5 years), gender, smoking status, diabetes status and hypertension status, each edentulous patient was paired with dentulous individuals suffering from various severity of periodontitis in the same cohort. Then, we selected 3 groups of patients (n=27 in each group) with no or mild periodontitis (mild group), moderate periodontitis (moderate group) and severe periodontitis (severe group). The whole unstimulated saliva was collected before the periodontal examination. Questionnaire survey and periodontal parameters, including plaque index (PLI), probing depth (PD), bleeding index (BI) and clinical attachment loss (CAL), were examined at mesial-buccal and distal-lingual sites of each tooth respectively. DNA was extracted from each sample of the salivary deposition. Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), Campylobacter rectus (Cr) and Prevotella nigrescens (Pn) were detected by using PCR method based on 16SrRNA. The prevalence and quantity of the pathogens under various severity of periodontitis were compared. Results: One or more periodontal pathogens could be detected from the 78% (21/27) of the salivary samples in edentulous group. Thereinto, the prevalences of the five periodontal pathogens were ranked as (from high to low): Cr [56% (15/27)], Tf [44% (12/27)], Pn [26% (7/27)], Pg [22% (6/27)] and Td [11% (3/27)]. All five pathogens' prevalences and Pg, Tf, Td and Pn's quantities showed statistical differences among the four groups. The numbers of detected bacterial species in the mild, moderate and severe groups were significantly higher than that in the edentulous group (P<0.01). Furthermore, the prevalences of the red complex in three dentulous groups [96% (26/27) in each group] were significantly higher than the edentulous group [48% (13/27)] (P<0.05). The proportions of the red complex among all five pathogens (83%) in moderate and severe groups were significantly higher than that in the edentulous group (37%) (P<0.01). Conclusions: All five periodontal pathogens could be detected in most of the saliva samples from edentulous individuals. Nevertheless, the prevalence and quantity were lower than dentulous individuals.
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Saliva , Pequim , Pré-Escolar , Estudos Transversais , HumanosRESUMO
Nasopharyngeal carcinoma (NPC) is an aggressive head and neck malignancy, and radiotherapy (with or without chemotherapy) is the primary treatment modality. Reliable tumour assessment during the treatment phase, which can portend the efficacy of radiotherapy and early identification of potential treatment failure in radioresistant disease, has been implicit for better cancer management. Technological advancement in the last decade has fostered the development of functional magnetic resonance imaging (fMRI) techniques into a promising tool for diagnostic and therapeutic assessments in head and neck cancer. Apart from conventional morphological assessment, early detection of the physiological environment by fMRI allows a more thorough investigation in monitoring tumour response. This article discusses the relevant fMRI utilities in NPC as an early prognostic and monitoring tool for treatment. Challenges and future developments of fMRI in radiation oncology are also discussed.
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Neoplasias de Cabeça e Pescoço , Neoplasias Nasofaríngeas , Meios de Contraste , Imagem de Difusão por Ressonância Magnética , Humanos , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância Magnética , Carcinoma Nasofaríngeo/diagnóstico por imagem , Carcinoma Nasofaríngeo/radioterapia , Neoplasias Nasofaríngeas/diagnóstico por imagem , Neoplasias Nasofaríngeas/radioterapia , PrognósticoRESUMO
BACKGROUND: The amplified in breast cancer 1 (AIB1) gene has been considered to play an oncogenic role in human cancers, but its clinical/prognostic significance in non-small-cell lung cancer (NSCLC) is still unclear. PATIENTS AND METHODS: The methods of immunohistochemistry and FISH were utilized to examine protein expression and amplification of AIB1 in 230 informative surgically resected NSCLCs and in 30 samples of normal lung tissues. RESULTS: Overexpression and amplification of AIB1 were found in 48.3% and 8.2% of NSCLCs, respectively. AIB1 overexpression was associated with AIB1 gene amplification and cell proliferation but not related to estrogen receptor (ER)-alpha, ER-beta, progesterone receptor or androgen receptor status. A positive correlation between AIB1 overexpression and an ascending pathologic node stage in lung adenocarcinoma (ADC) was observed (P = 0.043). Univariate survival analysis demonstrated a significant association of AIB1 overexpression with shortened patient survival, especially for those with stage III disease (P < 0.001). Importantly, AIB1 expression was evaluated as the most significant predictor for survival in multivariate analysis (hazards ratio = 2.069, P < 0.001). CONCLUSION: Overexpression of AIB1 might provide a selective advantage for lymph node metastasis of lung ADC and serve as a useful biomarker for poor prognosis for NSCLC patients.
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Carcinoma Pulmonar de Células não Pequenas/patologia , Neoplasias Pulmonares/patologia , Coativador 3 de Receptor Nuclear/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/cirurgia , Feminino , Humanos , Hibridização in Situ Fluorescente , Pulmão/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/cirurgia , Masculino , Pessoa de Meia-Idade , Prognóstico , Receptores Androgênicos/metabolismo , Receptores de Estrogênio/metabolismo , Receptores de Progesterona/metabolismo , Taxa de SobrevidaRESUMO
Clusterin (CLU) is expressed in a wide variety of human tissues and fluids. Overexpression of cytoplasmic clusterin (sCLU) has been implicated in cancer development and progression. The aim of the present study is to evaluate the association of sCLU overexpression with clinicopathological features of human gastric carcinomas (GC).We constructed a gastric cancer tissue microarray containing 173 primary gastric carcinomas and 70 paired non-neoplastic mucosa specimens. The expression of sCLU was studied by immunohistochemistry. The correlations between sCLU expression and clinicopathological features, p53 abnormality, as well as Ki67 activation were analyzed. Overexpressions of sCLU was detected in 28.5% (n=165) of primary GCs by immunohistochemical staining, but not in non-neoplastic mucosa. Clinical association study found that overexpression of sCLU was significantly correlated with lymph-node metastasis (p < 0.001), tumor invasion (p < 0.001) and TNM stage (p < 0.001). In Kaplan-Meier survival analysis, overexpression of sCLU was significantly correlated with unfavorable survival in advanced GCs (p < 0.03). Furthermore, the association of sCLU with abnormal expression of p53 was ascertained. These results suggested that overexpression of sCLU was involved in the progression of GC and it's oncogenic function might be associated with p53 abnormality. Overexpression of sCLU seems to be related with patient's shorter survival in late stage GC.
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Clusterina/fisiologia , Neoplasias Gástricas/patologia , Análise Serial de Tecidos/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Clusterina/análise , Citoplasma/química , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Estadiamento de Neoplasias , Prognóstico , Neoplasias Gástricas/química , Neoplasias Gástricas/mortalidade , Proteína Supressora de Tumor p53/análiseRESUMO
Members of the recently recognized SRC-1 family of transcriptional coactivators interact with steroid hormone receptors to enhance ligand-dependent transcription. AIB1, a member of the SRC-1 family, was cloned during a search on the long arm of chromosome 20 for genes whose expression and copy number were elevated in human breast cancers. AIB1 amplification and overexpression were observed in four of five estrogen receptor-positive breast and ovarian cancer cell lines. Subsequent evaluation of 105 unselected specimens of primary breast cancer found AIB1 amplification in approximately 10 percent and high expression in 64 percent of the primary tumors analyzed. AIB1 protein interacted with estrogen receptors in a ligand-dependent fashion, and transfection of AIB1 resulted in enhancement of estrogen-dependent transcription. These observations identify AIB1 as a nuclear receptor coactivator whose altered expression may contribute to development of steroid-dependent cancers.
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Neoplasias da Mama/genética , Amplificação de Genes , Neoplasias Hormônio-Dependentes/genética , Neoplasias Ovarianas/genética , Receptores de Estrogênio/metabolismo , Sequência de Aminoácidos , Mama/metabolismo , Neoplasias da Mama/metabolismo , Cromossomos Humanos Par 20 , Clonagem Molecular , Estradiol/metabolismo , Estradiol/farmacologia , Feminino , Dosagem de Genes , Regulação Neoplásica da Expressão Gênica , Histona Acetiltransferases , Humanos , Hibridização in Situ Fluorescente , Ligantes , Dados de Sequência Molecular , Neoplasias Hormônio-Dependentes/metabolismo , Coativador 1 de Receptor Nuclear , Coativador 2 de Receptor Nuclear , Neoplasias Ovarianas/metabolismo , Receptores de Estrogênio/genética , Fatores de Transcrição/genética , Ativação Transcricional , Transfecção , Células Tumorais CultivadasRESUMO
AIMS: To express and product a fluorescent antioxidant holo-alpha-phycocyanin (PC) of Spirulina platensis (Sp) with His-tag (rHHPC; recombinant holo-alpha-phycocyaninof Spirulina platensis with His-tag) in 5-l bench scale. METHODS AND RESULTS: A vector harbouring two cassettes was constructed: cpcA along with cpcE-cpcF in one cassette; ho1-pcyA in the other cassette. Lyases CpcE/F of Synechocystis sp. PCC6803 (S6) could catalyse the 82 site Cys in apo-alpha-PC of Sp linking with bilin chromophores, and rHHPC was biosynthesized in Escherichia coli BL21. The constant feeding mode was adopted, and transformant reached the biomass of rHHPC up to 0.55 g l(-1) broth in 5-litre bench scale. rHHPC was purified by Ni(2+) affinity column conveniently. The absorbance and the fluorescence emission spectra of rHHPC had lambda(max) at 621 and 650 nm, respectively. The IC(50) values of rHHPC were 277.5 +/- 25.8 microg ml(-1) against hydroxyl radicals and 20.8 +/- 2.2 microg ml(-1) against peroxyl radicals. CONCLUSIONS: Combinational biosynthesis of rHHPC was feasible, and the constant feeding mode was adopted to produce good yields of rHHPC. Fluorescent rHHPC with several unique qualitative and quantitative features was effective on scavenging hydroxyl and peroxyl radicals. SIGNIFICANCE AND IMPACT OF THE STUDY: A potent antioxidant rHHPC was co-expressed, produced and characterized for nutritional and pharmacological values, which would help to develop phycobiliproteins' applications in their fluorescent and biological activities.
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Ficocianina/biossíntese , Spirulina/metabolismo , Proteínas de Bactérias/química , Escherichia coli/genética , Escherichia coli/metabolismo , Vetores Genéticos , Radical Hidroxila/metabolismo , Liases/química , Mutagênese Insercional , Ficocianina/química , Ficocianina/genética , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Espectrometria de Fluorescência , Spirulina/genética , Synechocystis/genética , Synechocystis/metabolismoAssuntos
Biomarcadores Tumorais/análise , Carcinoma de Células Escamosas/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Carcinoma de Células Escamosas/patologia , Técnicas Citológicas , Detecção Precoce de Câncer , Feminino , Humanos , Imuno-Histoquímica/métodos , Neoplasias do Colo do Útero/patologia , Esfregaço Vaginal/métodosRESUMO
Esophageal squamous cell carcinoma (ESCC) is highly prevailing in Asia and it is ranked in the most aggressive squamous cell carcinomas. High-frequency loss of heterozygosity occurred in chromosome 14q11.2 in many tumors including ESCC, suggesting that one or more tumor-suppressor genes might exist within this region. In this study, we identified the tumor-suppressing role of DHRS2 (short-chain dehydrogenase/reductase family, member 2) at 14q11.2 in ESCCs. Downregulation of DHRS2 occurred in 30.8% of primary ESCC tumor tissues vs paired non-tumorous tissues. DHRS2 downregulation was associated significantly with ESCC invasion, lymph nodes metastasis and clinical staging (P<0.001). Survival analysis revealed that DHRS2 downregulation was significantly associated with worse outcome of patients with ESCC. In vitro and in vivo studies indicated that both DHRS2 variants could suppress cell proliferation and cell motility. Moreover, we demonstrated that DHRS2 could reduce reactive oxygen species and decrease nicotinamide adenine dinucleotide phosphate (oxidized/reduced), increase p53 stability and decrease Rb phosphorylation; it also decreased p38 mitogen-activated protein kinase phosphorylation and matrix metalloproteinase 2. In summary, these findings demonstrated that DHRS2 had an important part in ESCC development and progression.
Assuntos
Oxirredutases do Álcool/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/secundário , Movimento Celular , Proliferação de Células , Neoplasias Esofágicas/patologia , Regulação Neoplásica da Expressão Gênica , Proteínas Nucleares/metabolismo , Oxirredutases do Álcool/genética , Animais , Apoptose , Biomarcadores Tumorais/genética , Carbonil Redutase (NADPH) , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Estudos de Casos e Controles , Ciclo Celular , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/metabolismo , Feminino , Seguimentos , Humanos , Metástase Linfática , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Invasividade Neoplásica , Proteínas Nucleares/genética , Prognóstico , Taxa de Sobrevida , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
OBJECTIVE: To explore the role of LINC00963 in the pathogenesis of hepatocellular carcinoma and its underlying mechanisms. PATIENTS AND METHODS: The expression level of LINC00963 in 48 cases of hepatocellular carcinoma (HCC) tissues and paracancerous tissues were detected by quantitative Real-time (qRT-PCR). Survival analysis was carried out based on the expression level of LINC00963. The association between the expression level of LINC00963 and clinical characteristics of these subjects was analyzed by x2-test. The proliferation and cell cycle of HCC cells after transfection of LINC00963 overexpression plasmids were evaluated by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. RESULTS: The expression level of LINC00963 in HCC tissues was remarkably higher than that in paracancerous tissues, indicating a potential diagnostic significance of LINC00963. The progression-free -with the tumor size and TNM stage, but not with age, gender, histological type and lymph node metastasis. Overexpression of LINC00963 significantly enhanced the proliferation ability of HepG2 and HCC cells and prolonged their G0/G1 phase. Furthermore, the PI3K/AKT expression was increased after overexpression of LINC00963, while AKT siRNA effectively reversed the prolonged G0/G1 phase caused by LINC00963 overexpression. CONCLUSIONS: Our data revealed that LINC00963 was upregulated in HCC, which significantly extended the G0/G1 phase of HCC cells by activating PI3K/AKT pathway and promoting the proliferative ability of HCC cells. LINC00963 may be involved in the HCC development.