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1.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-941048

RESUMO

OBJECTIVE@#To investigate the effect of JAG1 on the malignant phenotype of triple-negative breast cancer (TNBC) and its role in angiogenesis in breast cancer microenvironment.@*METHODS@#The expressions of Notch molecules were detected in human TNBC 231 and 231B cells using RT-qPCR. Five female nude mice were inoculated with 231 cells and another 5 with 231B cells into the mammary fat pads, and 4-6 weeks later, the tumors were collected for immunohistochemical and immunofluorescence tests. 231 cells and 231B cells were treated with recombinant JAG (rJAG) protein and DAPT, respectively, and changes in their malignant phenotypes were assessed using CCK-8 assay, Hoechst 33258 staining, wound healing assay, Transwell chamber assay and endothelial cell adhesion assay. Western blotting was used to detect the changes in the expressions of proteins related with the malignant phenotypes of 231 and 231B cells. The effects of conditioned medium (CM) derived from untreated 231 and 231 B cells, rJAG1-treated 231 cells and DAPT-treated 231B cells on proliferation and tube formation ability of cultured human umbilical vein endothelial cells (HUVECs) were evaluated using CCK-8 assay and tube-forming assay.@*RESULTS@#The expression of JAG1 was higher in 231B cells than in 231 cells (P < 0.05). Tumor 231B showed higher expression of VEGFA and CD31. Compared with 231-Blank group, the migration, invasion and adhesion of 231 cells in 231-rJAG1 were significantly enhanced (P < 0.05). Protein levels of Twist1 and Snail increased (P < 0.01), anti-apoptotic protein Bcl-2 increased (P < 0.05), while DAPT inhibited the related phenomena and indicators of 231B. The 231-rJAG1-CM increased the cell number and tubule number of HUVEC (P < 0.05).@*CONCLUSION@#JAG1 may affect the malignant phenotype of TNBC and promote angiogenesis in the tumor microenvironment.


Assuntos
Animais , Feminino , Humanos , Camundongos , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Meios de Cultivo Condicionados , Células Endoteliais da Veia Umbilical Humana/metabolismo , Proteína Jagged-1/metabolismo , Camundongos Nus , Neovascularização Patológica/metabolismo , Inibidores da Agregação Plaquetária , Sincalida/metabolismo , Neoplasias de Mama Triplo Negativas/metabolismo , Microambiente Tumoral
2.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-775376

RESUMO

A new isobenzofuranone derivative has been isolated from Phlomis betonicoides by using various chromatographic techniques, including silica gel, Sephadex LH-20, MCI-gel resin and RP-HPLC. This compound was determined as 5-(3-hydroxypropyl)-2,2-dimethyl-2-furo[3,4-]chromen-7(9)-one (1) by NMR, MS, IR and UV spectroscopic data. Compound 1 showed potent antibacterial activity with an MIC₉₀ value of (58.4 ± 4.2) mg·L⁻¹ for methicillin resistant Staphylococcus aureus (MRSA) strain [levofloxacin as a control with MIC₉₀ value of (52.8±4.6) mg·L⁻¹].


Assuntos
Antibacterianos , Farmacologia , Benzofuranos , Farmacologia , Staphylococcus aureus Resistente à Meticilina , Testes de Sensibilidade Microbiana , Phlomis , Química , Compostos Fitoquímicos , Farmacologia
3.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-692249

RESUMO

To measure particle size distribution of phenols in mainstream cigarette smoke aerosol,the particles of cigarette smoke aerosol were divided into 12 stages using single channel smoking machine coupled electrical low-pressure impactor (ELPI) and collected by 12 polyester films.The collected particles were weighted and then analyzed by ultra-high performance liquid chromatography-fluorescence detection (UPLC-FLD) to determine the 14 phenols in the different size particles.The results showed that the aerosols collecting method had good stability with relative standard deviation (RSD) of collected particles mass less than 10%.The analyzing results of 14 phenols by UPLC-FLD showed that the linear correlation coefficients(R2) were greater than 0.9959,with detection limits were less than 1.2 ng/cig and recoveries were 80.1%-115.0%.The distributions of 14 phenols with respect to smoke aerosol particle size were investigated.The results indicated that except 4-ethyl-2-methoxy-phenol not detected,the other 13 phenols were detected in mainstream cigarette smoke aerosol.The content of 13 phenols appeared increasing at first and then decreasing with increase of the particle size which distributed in a pattern similar to that of particle mass.All of 13 phenols were present in higher amounts in the medium size particles (0.261-0.722 μm) with peak content in particles 0.431 μm.The distribution of concentrations (ratio of content to particle mass) of 13 phenols in different size particles was different.The concentrations of phenol and mono-substituted phenol appeared to first increase and then decrease with increasing smoke aerosol particle size and were higher in medium size particles (0.261-0.772 μm).The concentrations of benzenediol and mono-substituted benzenediol were uniformly distributed in medium size particles (0.144-1.166 μm),and the concentration of disubstituted phenol was uniform throughout the particles of varying sizes.

4.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-272632

RESUMO

<p><b>OBJECTIVE</b>To study the roles of DNA dependent protein kinase (DNA-PK)in silica-induced cell cycle changes and expressions of CyclinE and CDK2 in human embryo lung fibroblasts (HELF).</p><p><b>METHODS</b>The expressions of Ku80 and DNA-PKcs proteins were inhibited by siRNA plasmids, respectively. Flow cytometry was used to detect the distributions of cell cycle and western blot assay was used to determine the expression levels of CyclinE and CDK2 after cells were exposed to 200 microg/ml silica for 0, 3, 6, 12, 24 h.</p><p><b>RESULTS</b>The proportion of G1 phases in negative control cells decreased from 83.53% +/- 2.24% to 69.11% +/- 3.12% after exposure to silica; the proportion of G1 phases in H-Ku80 and H-PKcs cells exposed to silica decreased from 85.16% +/- 3.73% to 59.92% +/- 3.31% and from 75.06% +/- 2.23% to 58.32% +/- 1.35%, respectively (P < 0.05). The exposure to silica resulted in the increasing protein expression levels of CyclinE and CDK2 in negative control cells, and the expression levels of CyclinE were obviously suppressed in H-Ku80 and H-PKcs as compared with control cells. However, the expression level of CDK2 protein did not change significantly.</p><p><b>CONCLUSION</b>DNA-PK might play a role in silica-induced alternations of cell cycle and regulate silica-induced overexpression of CyclinE in human embryo lung fibroblasts.</p>


Assuntos
Humanos , Ciclo Celular , Células Cultivadas , Ciclina E , Metabolismo , Quinase 2 Dependente de Ciclina , Metabolismo , Proteína Quinase Ativada por DNA , Genética , Metabolismo , Fibroblastos , Biologia Celular , Metabolismo , Pulmão , Biologia Celular , Proteínas Nucleares , Genética , Metabolismo , Proteínas Oncogênicas , Metabolismo , Dióxido de Silício , Farmacologia
5.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-272608

RESUMO

<p><b>OBJECTIVE</b>To study the roles of Ku80/p53 pathway in silica-induced cell cycle changes in human embryo lung fibroblasts (HELF).</p><p><b>METHODS</b>Ku80 siRNA expression vectors were transfected into HELF by lipofectamine. Flow cytometry was used to detect the distributions of cell cycle and western blot assay was used to determine the expression level of Ku80, p53 and p21 proteins or the phosphorylation levels of p53-ser15 after cells were exposed to silica.</p><p><b>RESULTS</b>The expression levels of Ku80 protein increased in concentration-dependent and time-dependent manners after cells were exposed to silica. The proportion of G1 phases in H-NC cells (controls) decreased from 89.28% +/- 2.19% to 68.93% +/- 3.79% after exposure to silica, and the proportion of G1 phases in HELF cells (H-Ku80) decreased from 85.16% +/- 3.73% to 59.92% +/- 3.31% after exposure to silica (P<0.05). The expression levels of Ku80, p53 proteins or p21 proteins or phosphorylation level of p53-ser15 were obviously suppressed in H-Ku80, as compared with H-NC.</p><p><b>CONCLUSION</b>Ku80/p53 pathway plays a role in the cell cycle charges induced by silica in human embryo lung fibroblasts.</p>


Assuntos
Humanos , Antígenos Nucleares , Metabolismo , Ciclo Celular , Células Cultivadas , Inibidor de Quinase Dependente de Ciclina p21 , Metabolismo , Proteínas de Ligação a DNA , Metabolismo , Fibroblastos , Biologia Celular , Metabolismo , Citometria de Fluxo , Autoantígeno Ku , Pulmão , Biologia Celular , Metabolismo , Fosforilação , Quartzo , Toxicidade , Transdução de Sinais , Proteína Supressora de Tumor p53 , Metabolismo
6.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-269661

RESUMO

Tumors often have DNA repair defects, suggesting additional inhibition of other DNA repair pathways in tumors may lead to synthetic lethality. Accumulating data demonstrate that DNA repair-defective tumors, in particular homologous recombination (HR), are highly sensitive to DNA-damaging agents. Thus, HR-defective tumors exhibit potential vulnerability to the synthetic lethality approach, which may lead to new therapeutic strategies. It is well known that poly (adenosine diphosphate (ADP)-ribose) polymerase (PARP) inhibitors show the synthetically lethal effect in tumors defective in BRCA1 or BRCA2 genes encoded proteins that are required for efficient HR. In this review, we summarize the strategies of targeting DNA repair pathways and other DNA metabolic functions to cause synthetic lethality in HR-defective tumor cells.


Assuntos
Animais , Humanos , Antineoplásicos , Farmacologia , Neoplasias da Mama , Genética , Reparo do DNA , Genética , Regulação Neoplásica da Expressão Gênica , Genes Letais , Genética , Genes Supressores de Tumor , Genes cdc , Mutagênese , Inibidores de Poli(ADP-Ribose) Polimerases , Proteína Rad52 de Recombinação e Reparo de DNA , Recombinação Genética , Genética
7.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-282568

RESUMO

<p><b>OBJECTIVE</b>To investigate the roles of mitogen-activated protein kinases (MAPK) on silica-induced cell cycle changes.</p><p><b>METHODS</b>After cells were treated with 200 microg/ml silica, Western blot and Immunofluorescence assays were utilized to detect the expression of cyclin D1, CDK4 and E2F-4, Flow cytometry was used to detect cell cycle progression, the dominant negative mutants techniques were used to investigate silica-induced signal pathway and the effects of which in silica-induced cell cycle changes.</p><p><b>RESULTS</b>After cells were exposed to 200 microg/ml silica 24 h, the results of present study showed the proportion of cells in G1 phases was decreased. Silica-induced cell cycle alternation was markedly impaired by stable expression of a dominant negative mutants of ERK or JNK, but not p38. It was found that ERK and JNK were involved in silica-induced cyclin D1 and CDK4 overexpression and the decreased expression of E2F-4.</p><p><b>CONCLUSION</b>ERK and JNK, but not p38, mediated silica-induced cell cycle changes in human embryo lung fibroblasts.</p>


Assuntos
Humanos , Ciclo Celular , Divisão Celular , Células Cultivadas , Fibroblastos , Biologia Celular , Patologia , Sistema de Sinalização das MAP Quinases , Proteínas Quinases Ativadas por Mitógeno , Metabolismo , Quartzo , Toxicidade
8.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-282503

RESUMO

<p><b>OBJECTIVE</b>To investigate the roles of cyclin D1 and CDK4 in the cell cycle changes of human embryonic lung fibroblasts (HELFs) exposed to silica.</p><p><b>METHODS</b>HELFs were divided into 4 groups: control group, curcumin (20 µmol/L for 1 h) group, silica (200 µg/ml for 24 h) group and curcumin plus silica group, i.e. after exposure to 20 µmol/L curcumin for 1h, the HELFs were treated with 200 µg/ml silica for 24 h. Western blot and Immunofluorescence assays were utilized to detect the expression levels of cyclin D1, CDK4 and E2F1/4. Flow cytometry was used to detect the cell cycle progression, the RNA transfection technique was used to investigate the silica-induced signal pathway and the roles of which in silica-induced cell cycle changes.</p><p><b>RESULTS</b>The expression levels of cyclin D1 and CDK4 significantly increased and the expression level of E2F-4 decreased obviously, but the expression level of E2F-1 did not significantly change in silica group. The proportion of G1 phase cells obviously decreased and the proportion of S phase cells significantly increased in silica group, as compared with control group (P < 0.05). When suppressing the expression of cyclin D1 or CDK4, the proportions of cells in G1 phase in anti-D1 plus silica group and anti-K4 plus silica group did not obviously change, as compared with control group. When suppressing AP-1 activity, the cyclin D1 and CDK4 expression levels decreased and the E2F-4 expression level increased in curcumin plus silica group, as compared with silica group.</p><p><b>CONCLUSION</b>The results of present suggested that 200 µg/ml silica could induce the high expression of cyclin D1 and CDK4 and the low expression of E2F-4, resulting in the cell cycle changes by AP-1/cyclin D1 pathway in HELFs.</p>


Assuntos
Humanos , Ciclo Celular , Células Cultivadas , Ciclina D1 , Metabolismo , Quinase 4 Dependente de Ciclina , Metabolismo , Fator de Transcrição E2F4 , Metabolismo , Fibroblastos , Biologia Celular , Metabolismo , Fase G1 , Quartzo , Fator de Transcrição AP-1 , Metabolismo , Transfecção
9.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-671617

RESUMO

ObjectiveTo investigate the effects of mefformin on the levels of serum retinol-binding protein 4 (RBP-4),high sensitive C reactive protein (hs-CRP) and tumor necrosis factorα (TNF-α) in patients with impaired glucose tolerance(IGT) and metabolic syndrome(MS). MethodsSixty patients with IGT and MS were divided into mefformin treatment group (30 cases) and life-style intervention group (30 cases) by random digits table. Body mass index (BMI),the levels of HbAic, HOMA-IR, blood fat, RBP-4,hs-CRP, TNF- α were measured both before and 16 weeks after treatment in the two groups and compared.ResultsThe levels of HbA1c, HOMA-IR, RBP-4, hs-CRP and TNF- α were significantly lower in mefformin treatment group than those in life-style intervention group [(5.09 + 0.26 )% vs. (5.69 ± 0.49 )%, 2.95 ± 0.63vs. 3.49 ± 0.78, ( 18.69 ± 6.50) mg/L vs.(26.20 ± 6.97) mg/L, (2.37 ± 0.53) mg/L vs.(2.99 ± 0.57) mg/L,(9.49 ± 2.37) μ g/L vs. ( 14.33 ± 2.62) μ g/L] (P < 0.01 ). The results of multiple linear regression showed that correlations were found between the changes of RBP-4 and BMI,HOMA-IR,hs-CRP,TNF-α(β =0.284,0.506,0.274,0.230,P <0.01 ),and HOMA-IR was the most important limiting factor. Conclusions Mefformin can improve insulin sensitivity of the patients with IGT and MS, and depress the levels of RBP-4,hs-CRP and TNF- α. Meanwhile mefformin has anti-inflammatory effect.

10.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-275741

RESUMO

<p><b>OBJECTIVE</b>To study the role of Phosphatidylinositol 3 kinase (PI3K) in silica-induced DNA double strand break repair in human embryo lung fibroblasts (HELF).</p><p><b>METHODS</b>Control HELF cells and DN-Deltap85 (HELF transfected with Dominant negative mutant of PI3K) were treated with 200 microg/ml silica for different times. The expression levels of phosphor-H2AX (H2AX), Ku70, Ku80 and DNA-PKcs were determined by Western blot. Furthermore, DNA double strand breaks were measured by neutral comet assay after cells were treated with 200 microg/ml silica for 0, 12 and 24 h.</p><p><b>RESULTS</b>After treatment with 200 microg/ml silica for different times, the levels of H2AX were increased in a time-dependent manner and the expression levels of H2AX were obviously suppressed in DN-Deltap85 compared with control cells. The levels of Ku70 and Ku80 were also significantly suppressed in DN-Deltap85 (0.37 +/- 0.14, 0.55 +/- 0.17) compared with control cells (0.58 +/- 0.09, 0.95 +/- 0.21) after treatment with 200 microg/ml silica for 12 h (P < 0.05). Both the percentage of tail DNA in HELF and DN-Deltap85 increased significantly at 12 h (9.78 +/- 1.15, 11.79 +/- 4.90) compared with groups without treatment with silica (2.40 +/- 0.69, 3.31 +/- 1.35) and then decreased at 24 h (4.19 +/- 0.47, 7.58 +/- 4.32), but only the decrease of HELF at 24 h was significant compared with HELF at 12 h (P < 0.05). DNA repair competence of HELF was 75.74% and that of DN-Deltap85 declined to 49.64%.</p><p><b>CONCLUSION</b>Silica dust can induce DNA double strand breaks in human embryo lung fibroblasts. PI3K might play a role in silica-induced DNA double strand break repair by regulating the expression levels of Ku70 and Ku80.</p>


Assuntos
Humanos , Antígenos Nucleares , Metabolismo , Proteínas de Ligação ao Cálcio , Metabolismo , Células Cultivadas , Ensaio Cometa , Quebras de DNA de Cadeia Dupla , Dano ao DNA , Reparo do DNA , Proteínas de Ligação a DNA , Metabolismo , Fibroblastos , Histonas , Metabolismo , Autoantígeno Ku , Pulmão , Biologia Celular , Fosfatidilinositol 3-Quinase , Metabolismo , Dióxido de Silício , Toxicidade
11.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-275740

RESUMO

<p><b>OBJECTIVE</b>To study the role of p53 in silica-induced cell cycle alternation and DNA double strand breaks repair in human embryo lung fibroblasts (HELF).</p><p><b>METHODS</b>Neutral comet assay was applied to detect silica-induced DNA double strand breaks. According to the neutral comet experimental result, the DNA repair competence was calculated. The expression levels and phosphorylation of protein in HELF were determined by Western blot. Cell cycle changes were identified by flow cytometry in HELF.</p><p><b>RESULTS</b>After treatment with 200 microg/ml silica for different times (0, 1, 2, 6, 12 and 24 h), the expression levels and phosphorylation of p53 increased in a time-dependent manner, reaching maximum at 12 h and then decreasing at 24 h. After treatment with 0, 25, 50, 100, 200, 300 and 400 microg/ml silica for 12 h, the expression levels and phosphorylation of p53 increased in concentration-dependent manner. After p53 expression was inhibited, silica-induced DNA damage repair competence was markedly increased (DRC = 87.68%), compared with the negative control cell induced by silica (DRC = 57.19%). Silica increased the percentage of S phase (31.8 +/- 1.1)% compared with the controls (24.3 +/- 3.8)% (P < 0.05). When p53 expression was inhibited, the number of S phase cells was significantly increased, (41.4 +/- 0.6)% compared with the controls (25.4 +/- 1.9)% (P < 0.05).</p><p><b>CONCLUSION</b>The silica dramatically increases the expression levels and phosphorylation of p53. The increased expression of p53 mediates silica-induced cell cycle change and inhibits silica-induced DNA double strand breaks repair.</p>


Assuntos
Humanos , Ciclo Celular , Linhagem Celular , Ensaio Cometa , Quebras de DNA de Cadeia Dupla , Dano ao DNA , Reparo do DNA , Fibroblastos , Biologia Celular , Metabolismo , Pulmão , Biologia Celular , Dióxido de Silício , Toxicidade , Proteína Supressora de Tumor p53 , Metabolismo
12.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-347259

RESUMO

<p><b>OBJECTIVE</b>To study the role of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) in silica-induced DNA double-strand break repair in human embryo lung fibroblasts (HELF).</p><p><b>METHODS</b>Two stable transfectants, HELF transfected with DNA-PKcs siRNA (HELF-PKcs) and with negative control siRNA (HELF-NC), were established. HELF cells were treated with 0, 25, 50, 100, 200, 300 and 400 microg/ml silica for 12 h and with 200 microg/ml silica for different times (0, 1, 2, 6, 12 and 24 h). HELF-PKcs and HELF-NC were treated with 200 microg/ml silica for 0, 12 and 24 h. The expression levels of DNA-PKcs and phosphor-H2AX (H2AX) were determined by Western blot. DNA double strand breaks were measured by neutral comet assay.</p><p><b>RESULTS</b>After treatment with different doses of silica for 12 h, the levels of H2AX and the percentages of tail DNA increased in concentration-dependent manner. After treatment with 200 microg/ml silica for different times, the levels of H2AX increased in a time-dependent manner. The percentages of tail DNA increased significantly at 6 h, and reaching maximum at 12 h and then decreasing at 24 h. The expression level of DNA-PKcs was suppressed in HELF-PKcs. After treatment with silica at 12 h, the level of H2AX was lower in HELF-PKcs than in HELF-NC, and the percentages of tail DNA increased obviously in both HELF-PKcs and HELF-NC compared with non-treated cells, but no significant difference was found in the percentages of tail DNA between them. The percentages of tail DNA decreased markedly in silica-treated HELF-NC and was significantly lower than in HELF-PKcs at 24 h (P < 0.05).</p><p><b>CONCLUSION</b>Silica can induce DNA double strand breaks in human embryo lung fibroblasts. DNA-PKcs might play a major role in silica-induced DNA double strand break repair. Silica-induced histone H2AX phosphorylation was dependent on DNA-PKcs.</p>


Assuntos
Humanos , Linhagem Celular , Quebras de DNA de Cadeia Dupla , Reparo do DNA , Proteína Quinase Ativada por DNA , Genética , Metabolismo , Fibroblastos , Fisiologia , Histonas , Metabolismo , Fosforilação , Dióxido de Silício , Farmacologia , Transfecção
13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-304057

RESUMO

<p><b>OBJECTIVE</b>To investigate the roles of activated protein 1 (AP-1) in cell cycle changes on human embryo lung fibroblasts (HELF) induced by benzo (a) pyrene [B (a) P], and relationships between AP-1 and cyclin D1/CDK4-E2F-1/4.</p><p><b>METHODS</b>Cells transfected with AP-1 luciferase reporter plasmid (AP-H) were cultured with serum-free RPMI1640 for 48 h, and treated with 2 micromol/L B (a) P for 24 h. AP-1 relative activity was detected by luciferase assay. Changes of cell cycle and the expression of cyclin D1, CDK4 and E2F-1/4 were checked using the flow cytometer and Western blot assay.</p><p><b>RESULTS</b>After B (a) P was treated for 24 h, the ratio of G1 phase cells (71 +/- 2)% was decreased to (48 +/- 3)% (P < 0.05), and an increase was observed in the ratio of S phase. AP-1 activity and cyclin D1/E2F-1 expression were increased significantly, but CDK4/E2F-4 expression did not change after B (a) P treatment. When AP-1 activity was inhibited by curcumin, decreases of G1 phase in response to B (a) P treatment were blocked, and overexpression of cyclin D1/E2F-1 was attenuated, but CDK4/E2F-4 expression was not changed significantly.</p><p><b>CONCLUSION</b>AP-1 is involved in B (a) P induced cell cycle changes, and is the upstream signals of cyclin D1/E2F-1, but not CDK4/E2F-4.</p>


Assuntos
Humanos , Benzo(a)pireno , Toxicidade , Ciclo Celular , Células Cultivadas , Ciclina D1 , Metabolismo , Quinase 4 Dependente de Ciclina , Metabolismo , Fator de Transcrição E2F1 , Metabolismo , Fator de Transcrição E2F4 , Metabolismo , Fibroblastos , Biologia Celular , Metabolismo , Fator de Transcrição AP-1 , Genética , Metabolismo , Transfecção
14.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-352468

RESUMO

<p><b>OBJECTIVE</b>To investigate the roles of p53 in cell cycle changes on human embryo lung fibroblasts (HELF) induced by benzo(a) pyrene[ B(a) P], and relationships between p53 and p21, E2F-1.</p><p><b>METHODS</b>Cells transfected with p53 siRNA plasmid (p53-H) and CMV vector (HELF/CMV) were cultured with serum-free R/MINI-1640 for 48 hours, then treated with 2 micromol/L B(a)P for 24 hours. Flow cytometry assay was used for detecting the cell cycle alteration after being exposed to B(a)P. Changes of p53 and p21 expressions were checked using Western blot assay, and the cytoplasmic and nuclear extraction was used to observe the subcellular localizations of p53 and p21. The immunofluorescence assay was used to check changes of E2F-1 expression and the distribution of E2F-1 in nuclear and cytoplasm after exposed to B (a)P. p53siRNA plasmid and the chemical inhibitor of p53 [pifithrin-alpha (PFT)] were used to observe effects of p53 in B(a)P induced cell cycle changes and the relationships of p53 and p21, E2F-1.</p><p><b>RESULTS</b>After 2 micromol/L B(a)P exposure, the ratio of G1 phase cells (71 +/- 5)% was decreased to (39 +/- 4)% (P < 0.05). p53, p21 and E2F-1 expressions were increased significantly, and over expressed proteins were mostly located in nuclear after B(a)P treatment. When p53 expression was inhibited by p53 siRNA or PFT, the decreases of G1 phase in response to B(a)P treatment still existed, and over expression of p21 induced by B(a)P was attenuated, especially in nuclear, but E2F-1 over expression was not changed significantly.</p><p><b>CONCLUSION</b>B(a)P could induce cell cycle changes through p53 independent pathways. And p53 could regulate p21 expression positively, but not E2F-1.</p>


Assuntos
Humanos , Benzo(a)pireno , Toxicidade , Ciclo Celular , Células Cultivadas , Inibidor de Quinase Dependente de Ciclina p21 , Dano ao DNA , Fator de Transcrição E2F1 , Fibroblastos , Citometria de Fluxo , Expressão Gênica , Pulmão , Biologia Celular , Metabolismo , Mecanotransdução Celular , RNA Mensageiro , Genética , Proteína Supressora de Tumor p53 , Genética , Metabolismo
15.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-338907

RESUMO

<p><b>OBJECTIVE</b>To investigate the alteration of activator protein-1 (AP-1) luciferase activity in human embryo lung fibroblasts (HELF) after exposed to silica, and the role of mitogen activated protein kinase (MAPK)/AP-1 pathway on silica-induced cell cycle changes.</p><p><b>METHODS</b>After HELF cells were treated with 200 microg/ml silica, immunofluorescence assays were employed to detect the translocation and the phosphorylation level of extracellular signal-regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK), flow cytometry was used to detect the distributions of cell cycle, the dominant negative mutant of ERK, JNK and p38 were applied to detect the upstream or downstream relationship of signaling pathways.</p><p><b>RESULTS</b>After HELF-AP-1 cells were exposed to 200 microg/ml silica 6, 12, 24 h respectively, silica exposure lead to AP-1 activation in a time-dependent manner, inducing significant AP-1 activation at 6 h, reaching a maximum activation at 12 h, and having a little decrease at 24 h. After silica exposure 1 h, phosphorylation level of ERK and JNK increased mainly in cytoplasm, however, after exposure 2 h, they translocated to nucleus. The proportion of cells in G1 phases was decreased from (63.80 +/- 9.57)% to (32.23 +/- 7.22)%, and the proportion of cells in S phases was increased from (35.17 +/- 10.33)% to (66.00 +/- 8.07)% after exposed to silica 24 h. Curcumin, a chemical inhibitor of AP-1, impaired the decrease of cells in G1 phases. Furthermore we found expression of dominant-negative mutant of ERK and JNK impaired silica-induced AP-1 activation, whereas, dominant-negative mutant of p38 did not show the effect.</p><p><b>CONCLUSION</b>These result suggested that 200 microg/ml silica exposure can induce AP-1 activation, induce cell cycle changes through ERK, JNK/AP-1-dependent pathway.</p>


Assuntos
Humanos , Ciclo Celular , Células Cultivadas , MAP Quinases Reguladas por Sinal Extracelular , Metabolismo , Fibroblastos , Biologia Celular , Metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno , Metabolismo , Pulmão , Biologia Celular , Quartzo , Farmacologia , Transdução de Sinais , Fator de Transcrição AP-1 , Metabolismo
16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-290236

RESUMO

<p><b>OBJECTIVE</b>To explore the relation of body mass index (BMI) and waist circumference (WC) of the aged with clustering of other cardiovascular risk factors.</p><p><b>METHODS</b>Total 654 old people were checked up, and the systolic blood press (SBP), diastolic blood press (DBP), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and fasting plasma glucose (FPG) of them were investigated according to different BMI and WC, then the other cardiovascular risk factors analysis was carried out on the detection rates of SBP, DBP, TC, HDL-C, FPG level and high blood pressure, high TC, high TG. The clustering of the other cardiovascular risk factors was analyzed.</p><p><b>RESULTS</b>The rising of BMI and WC was regular cardiovascular risk factor in the aged. The average level of SBP, DBP, TC, TG, and FPG of the aged was obviously rising along with BMI and WC increasing, meanwhile the HDL-C was obviously decreased, and the detection rates of high blood pressure, high TC, TG and diabetes mellitus were obviously increased. When the BMI was 24 kg/m(2)-27.9 kg/m2, the detection rates of high blood pressure, high TC, TG and diabetes mellitus were 59.74%, 3.89%, 28.57%, 10.06% respectively, and their risks of suffering high blood pressure, high TC and TG were 1.65, 1.88, 1.85 times of those with normal BMI. When the BMI > or = 28.0 kg/m2, the detection rates of high blood pressure, high TC, TG and diabetes mellitus were 83.05%, 5.08%, 35.59%, 15.25% respectively, and their risks of suffering high blood pressure, high TC and TG were 5.44, 2.60, 2.98 times of those with normal BMI. When the man WC > or = 85 cm or women WC > or = 80 cm, the detection rates of high blood pressure, high TC, TG and diabetes mellitus were 66.15.%, 4.47%, 29.57%, 10.12%, and their risks of suffering high blood pressure, high TC and TG were 3.52, 6.51, 1.68 times of those with normal BMI. The rate of the aged with several cardiovascular risk factors was significantly increased for those with BMI > or = 24.0 kg/m2 or man WC > or = 85 cm or women WC > or = 80 cm.</p><p><b>CONCLUSION</b>The rising of BMI and WC should be important factor for the clustering of cardiovascular risk factors in the aged.</p>


Assuntos
Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores Etários , Pressão Sanguínea , Índice de Massa Corporal , Doenças Cardiovasculares , Sangue , Colesterol , Sangue , HDL-Colesterol , Sangue , Obesidade , Epidemiologia , Fatores de Risco , Triglicerídeos , Sangue , Circunferência da Cintura
17.
Chinese Journal of Epidemiology ; (12): 697-699, 2003.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-246453

RESUMO

<p><b>OBJECTIVE</b>To evaluate the immuno-effects of hepatitis B (HB) vaccination in adults.</p><p><b>METHODS</b>Five groups were sampled by means of cluster sampling, and serum HBsAg, anti-HBs and anti-HBc were tested in every group at people aged from 18 to 50. Recombinant HB vaccine was injected to the ones that HBsAg, anti-HBs and anti-HBc were all negative. Concentration of anti-HBs in serum was tested after one year and three years of vaccination. Immuno-effects of recombinant HB vaccination in adults at different ages and between sexes, were then calculated.</p><p><b>RESULTS</b>Good immuno-effects of recombinant HB vaccination in adults were noticed. After one year and three years of vaccination with 5 micro g recombinant HB vaccine, the anti-HBs positive rates were 82.76%, 70.77% while the serum concentrations of anti-HBs were 55.91 mIU/ml and 35.41 mIU/ml respectively. When 10 micro g was used, the concentrations were 83.74%, 72.22%, 56.89 mIU/ml and 30.29 mIU/ml respectively. The effects did not show significant differences between different doses on 10 micro g and of 5 micro g. Concentration of anti-HBs reduced when time went by. The factors such as age and sex influenced the effects of immunity on recombinant HB vaccination.</p><p><b>CONCLUSION</b>Good immunity could be obtained when recombinant hepatitis B was vaccinated in vulnerable population aged 18 to 50.</p>


Assuntos
Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Hepatite B , Anticorpos Anti-Hepatite B , Sangue , Antígenos do Núcleo do Vírus da Hepatite B , Alergia e Imunologia , Antígenos de Superfície da Hepatite B , Alergia e Imunologia , Vacinas contra Hepatite B , Alergia e Imunologia , Vacinação , Vacinas Sintéticas , Alergia e Imunologia
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