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Objective To investigate the effect of decitabine (DAC) on human acute myeloid leukemia (AML) cell line HL-60 and the regulating of natural killer (NK) cell activating receptor (NKG2D) ligands(NKG2DL), and to detect the molecular mechanism of JAK-STAT3-SOCS signaling pathway. Methods The effect of DAC on the proliferation of HL-60 was detected by using CCK-8 assay. The cell apoptosis was analyzed by using Annexin-V/PI double standard method. The expressions of receptor NKG2DL including MICA/B and ULBPs in HL-60 cells were detected by using flow cytometry (FCM). The killing activity of NK cells was analyzed by using carboxy fluorescein diacetate succinimidyl ester (CFSE). The expressions of JAK/STAT3 signaling pathway or molecules including STAT3, its upstream kinases JAK1, JAK2 and the negative regulator of STAT3,SOCS-1,SOCS-3 were examined by Western blot.Methylation level of the SOCS-1,SOCS-3 gene after the treatment of DAC was analyzed by using methylation-sensitive high resolution melting(MS-HRM). Results There was an obvious inhibitory effect of DAC on HL-60 cells. The cell viability of HL-60 treated with 0.2, 0.5, and 1.0 μmol/L DAC for 48 h was decreased by (25±11) %, (39±8) % and (50±7)%(P<0.01)respectively compared with those cells without DAC treatment.The incidence of apoptosis was (24.77±7.50) %, (27.10±4.48) % and (30.53±3.93) % after DAC treatment for 48h respectively, which were higher than that of untreated cells[(3.11±0.50)%](P<0.01).DAC induced a significant up-regulation of MICA/B, ULBP-1, ULBP-3 in HL-60 cells, and enhanced the sensitivity of HL-60 cells to NK cytotoxicity. Western blot results showed that a down-regulating expression of STAT3 and JAK1, JAK2 protein was detected, in addition to the phosphor-STAT3 and phosphor-JAKs in HL-60 cells after DAC treatment, but the expressions of SOCS-1 and SOCS-3 protein were increased. HRM results showed that DAC could inhibit the methylation of SOCS-3 gene. Conclusion DAC can inhibit the proliferation of HL-60 cells, upregulate the expression of NKG2DL and enhance the cytotoxicity of NK targeted to HL-60 cells, which might be related to the activity regulation of intracellular JAK-STAT3-SOCS signaling pathway.
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Objective To investigate the effect of decitabine (DAC) on human acute myeloid leukemia (AML) cell line HL-60 and the regulating of natural killer (NK) cell activating receptor (NKG2D) ligands(NKG2DL), and to detect the molecular mechanism of JAK-STAT3-SOCS signaling pathway. Methods The effect of DAC on the proliferation of HL-60 was detected by using CCK-8 assay. The cell apoptosis was analyzed by using Annexin-V/PI double standard method. The expressions of receptor NKG2DL including MICA/B and ULBPs in HL-60 cells were detected by using flow cytometry (FCM). The killing activity of NK cells was analyzed by using carboxy fluorescein diacetate succinimidyl ester (CFSE). The expressions of JAK/STAT3 signaling pathway or molecules including STAT3, its upstream kinases JAK1, JAK2 and the negative regulator of STAT3,SOCS-1,SOCS-3 were examined by Western blot.Methylation level of the SOCS-1,SOCS-3 gene after the treatment of DAC was analyzed by using methylation-sensitive high resolution melting(MS-HRM). Results There was an obvious inhibitory effect of DAC on HL-60 cells. The cell viability of HL-60 treated with 0.2, 0.5, and 1.0 μmol/L DAC for 48 h was decreased by (25±11) %, (39±8) % and (50±7)%(P<0.01)respectively compared with those cells without DAC treatment.The incidence of apoptosis was (24.77±7.50) %, (27.10±4.48) % and (30.53±3.93) % after DAC treatment for 48h respectively, which were higher than that of untreated cells[(3.11±0.50)%](P<0.01).DAC induced a significant up-regulation of MICA/B, ULBP-1, ULBP-3 in HL-60 cells, and enhanced the sensitivity of HL-60 cells to NK cytotoxicity. Western blot results showed that a down-regulating expression of STAT3 and JAK1, JAK2 protein was detected, in addition to the phosphor-STAT3 and phosphor-JAKs in HL-60 cells after DAC treatment, but the expressions of SOCS-1 and SOCS-3 protein were increased. HRM results showed that DAC could inhibit the methylation of SOCS-3 gene. Conclusion DAC can inhibit the proliferation of HL-60 cells, upregulate the expression of NKG2DL and enhance the cytotoxicity of NK targeted to HL-60 cells, which might be related to the activity regulation of intracellular JAK-STAT3-SOCS signaling pathway.
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Endemic dental fluorosis has been reported in some regions of the world. China seemed to have high prevalence of endemic dental fluorosis, especially in southwest China. It is now most likely that excessive fluoride intake during enamel development play a key role in the pathogenesis of dental fluorosis. However, excessive intake of fluoride-induced cellular and molecular mechanisms of dental fluorosis are not entirely conclusive. Scholars at home and abroad have made a lot of research on pathogenesis of enamel fluorosis by using various experimental techniques. More recent studies mainly suggest that endoplasmic reticulum stress and calcium overload-associated apoptotic pathway may participate in fluoride excess-evoked pathogenesis of dental fluorosis. Furthermore, the functional changes of enamel matrix protein and protease activity may be involved in the pathological event. This paper summarized the recent research progress on this topic.
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A lot of clinical trials proved that enhanced external counterpulsation (EECP) is a low-cost ,non-invasive , safe and effective method treating coronary heart disease .The present article made an overview on possible mecha-nism of EECP treating coronary heart disease .
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Pancreatic cancer normally is lethal and difficult to treat.Recently,although chemotherapy drugs such as gemcitabine for the treatment achieved a certain effect, but the survival rate has not improved significantly. Over the past decade, a large number of studies that aimed to implicated in pancreatic tumor gemcitabine metabolism and resistance mechanisms. Recent data show that it is a great value for predicting efficacy in the treatment of pancreatic cancer with gemcitabine by detecting pancreatic cancer -related molecules and genes, especially in the field of individual therapy. This review describes advances in the studies of related molecules and genes affecting the efficacy of gemcitabine for pancreatic cancer.
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Objective To study the effect of 2-methoxyestradiol (2-ME) on proliferation,apoptosis and its mechanism.Methods K562 cells were treated with 2-ME of different concentrations and time in vitro.Cell apoptosis rate was measured by flow cytometry(FCM).Activity of NF-Kappa B in nucleus was detected by electrophoretic molility shift assay(EMSA).Results After treatment with 2-ME,K562 cells apoptotic rate increased significantly.After treatment with 4μmol/L 2-ME for 24h、36h、48h,the activity of Caspase-3 and Caspase-9 were significantly higher and activity of NF-Kappa B in nucleus was significantly lower.Conclusion The present study showed that 2-ME induced apoptosis of K562 cells via active Caspase-3 and Caspase-9 and inhibit the activity of NF-kappa B in nucleus.This study provided useful experimental data for clinical application of 2-ME.
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Germinal center is a site of B cell maturation and development, in which B cell activation and differentiation was regulated by inherent complexity signal and transcription factors networks. Under the normal condition, the interaction between these transcription factors maintain dynamic stability of B cell in germinal center. When cytogenetic abnormalities or viral infection appear, a variety of.abnormal expression of transcription factors induced the imbalance of B cell signal networks regulation. manifested as abnormal B cell proliferation, apoptosis, and differentiation was blocked. B cell malignancies were often associated with genetic change of B cell and abnormal expression of transcription factors.
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Objective To investigate the relationship between the fluorouracil pathway gene and effect of chemotherapy in advanced gastric cancer after surgery. Methods 52 postoperative patients with advanced gastric cancer using FOLFOX4 6-cycles combined chemotherapy were collected to set up the database. The expression of thymidine synthase (TS), dihydropyrimidine dehydrogenase (DPD) and orotate phosphoribosyltransferase(OPRT) in tumor tissue and adjacent non-tumor tissue of 52 patients with advanced gastric cancer were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The influence of fluorouracil pathway gene on chemotherapy was investigated. Results The TS-mRNA level in tumor tissue was significantly higher than non-tumor tissue (0.92±0.28 vs 0.71±0.30) (t = 3.730, P =0.001).OPRT-mRNA level in tumor tissue was positively correlated with the non-tumor tissue (r =0.45, P =0.001). No correlations were observed among other gene expressions. Patients whose high OPRT-mRNA gene expression in their tumors and non-tumor tissue showed an obviously better survival (t = 3.036, P =0.003;t = 2.713, P =0.009). Patients with low DPD-mRNA gene expression survived longer than those with high DPD-mRNA gene expression in tumor tissue with statistical significance(t = 2.708, P =0.009), whereas prolonged survival was observed in patients with high DPD-mRNA gene expression in non-tumor tissue (t = 2.616, P =0.012).Conclusion There is close relationships between chemotherapy in advanced gastric cancer and the expression of DPD-mRNA, OPRT-mRNA;while the expression of TS-mRNA have no relation with the survival time.
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Mesenchymal stem cells (MSCs) not only exhibit a marked tropism for tumors,but also exert antitumorigenic activity or as an emerging vectors to express foreign genes continuously and efficiently within the tumor microenvironment,suppress tumor's growth and metastasis.Conversely,MSCs can accelerate tumor development through promoting tumor stroma remodeling and tumor vascular formation,inhibiting tumor local immune and changing tumor phenotype.The reason for this discrepancy may be attributable to different sources of MSCs,the heterogeneity of MSCs,differences in tumor microenviroment,or another factor that is not yet appreciated.
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In addition to kill tumor cells through direct cytotoxicity, the immunomodulatory activity is another facet of chemotherapy. The major molecular mechanisms of immunogenicity of cytotoxic drugs are elimination of CD_4~+ CD_(25)~+ regulatory T cells or Myeloid-Derived Suppressor Cells, and sensitization of tumor cells to immune effectors through modulation of death receptor expression. These studies provide scientific evidence for combinatorial treatments including vaccines and Traditional Chinese Medicine for cancer therapy.
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Objective: To explore the mechanism of damaging effect of advanced glycation end products (AGEs) on human umbilical vein endothelial cell (HUVEC) in vitro. Methods: The HUVEC was incubated with exogenous AGEs for 12 h, and ergamine (Hi) and catalase (CAT) were used as control. The activity, monolayer permeability, apoptosis rate, biochemical indexes and morphous changes were detected in HUVEC. Results: The activities of HUVEC were dose-dependently reduced by exogenous AGEs(40 mg/L, 120 mg/L and 160 mg/L), meanwhile the monolayer permeability, the malondialdehyde (MDA) content and apoptosis rate were increased，the activity of superoxide dismutase（SOD）was decreased（P < 0.05, respectively). There was no significant difference in damaging effect between exogenous AGEs and Hi(P ＞ 0.05). The damaging effect of exogenous AGEs was obviously inhibited by CAT in HUVEC. Conclusion: Exogenous AGEs induced the damaging effect on vascular endothelial cells, which may be related to the oxidative stress.
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Many studies indicate that macrophage migration inhibition factor(MIF)is over-expressed in tumor cells,and is involved in the carcinogenesis and tumor development by multiple methods and ways.The complicated molecular mechanisms are not quite clear,and the studies about MIF in digestive tumors,especially in hepatic cell carcinoma become more and more.
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Objective To investigate the anti-inflammatory effect of proanthocyanidins and its mechanisms.Methods Inflammation models such as dimethylbenzene-indueed ear swelling and carrageenan-induced hind paw edema in mice and rats were prepared.The contents of PGE2 in exudate from edema paws of rats were measured by ultraviolet spectrophotography and the protein expression of COX-2 in edema paws of rats by Western-blot and immunohistoehemistry(IHC)assay.Results Pro-anthocyanidins remarkably inhibited the ear edema induced by dimethylbenzene in mice at the dose of10 mg/kg and 20 mg/kg;paw edema of rats induced with carrageenan was significantly inhibited byproanthocyanidins 5,20 mg/kg ip from 2 to 5 h;proanthoeyanidins 5,20 mg/kg ip reduced the contents of PGE2 in exudate from edema paws of rats induced by carrageenan;proanthocyanidins 5,20 mg/kg ip inhibited the protein expression of COX-2.Conclusion Proanthocyanidins has an anti-inflammatory effect in vivo which may be related to inhibition of protein expression of COX-2 and downregutation of PGE2 biosynthesis.
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Objective To investigate the antibacterial molecular mechanism of Traditional Chinese Medicine Coptis rhizome against Yersinia pestis(Y.pestis).Methods The method based on whole genome DNA micrnarray of Y.pestis was used.The minimal inhibition concentration(MIC)of berberine to Y.pestis was determined with liquid dilution method.Then gene expression profile of Y.pestis was performed after exposed to berberine at the concentration of 10×MIC for 30 minutes.Total RNA extracted and purified from Y.pestis and reverse-transcribed to cDNA,then labeled by Cy-dye.Finally,the labeled probes were hybridized to the microarray and the results were obtained by a laser scanner and analyzed by the SAM software.Results The gene expression profile data revealed that the response of Y.pestis to berberine was a global phenomenon.A total of 360 genes changed significantly.Among them,333 genes were up-regulated,27 down-regulated.These differentially expressed genes were further classified into 24 different functional categories based on the genomie annotation of Y.pestis CO92,in which the number of mainly related genes were 83,75 and 48,including cell envelop,unkown,transport/binding proteins functions.The 40 genes related to the metabolism were upregulated,which was a remarkable change.Conclusion Our results have revealed the general gene expression changes of Y.pestis in response to berberine and demonstrated the antibacterial molecular mechanism of the Coptis rhizome.The major mechanism of Y.pestis in response to berberine is the upregulation of genes related to the metabolism.
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O principal determinante da nefropatia diabética é a hiperglicemia, mas hipertensão e fatores genéticos também estão envolvidos. O glomérulo é o foco de lesão, onde proliferação celular mesangial e produção excessiva de matriz extracelular decorrem do aumento da glicose intracelular, por excesso de glicose extracelular e hiperexpressão de GLUT1. Seguem-se aumento do fluxo pela via dos polióis, estresse oxidativo intracelular, produção intracelular aumentada de produtos avançados da glicação não enzimática (AGEs), ativação da via da PKC, aumento da atividade da via das hexosaminas e ativação de TGF-beta1. Altas concentrações de glicose também aumentam angiotensina II (AII) nas células mesangiais por aumento intracelular da atividade da renina (ações intrácrinas, mediando efeitos proliferativos e inflamatórios diretamente). Portanto, glicose e AII exercem efeitos proliferativos celulares e de matriz extracelular nas células mesangiais, utilizando vias de transdução de sinais semelhantes, que levam a aumento de TGF-beta1. Nesse estudo são revisadas as vias que sinalizam os efeitos da glicose e AII nas células mesangiais em causar os eventos-chaves relacionados à gênese da glomerulopatia diabética. As alterações das vias de sinalização implicadas na glomerulopatia, aqui revisadas, suportam dados de estudos observacionais/ensaios clínicos, onde controle metabólico e anti-hipertensivo, especificamente com inibidores do sistema renina-angiotensina, têm-se mostrado importantes - e aditivos - na prevenção do início e progressão da nefropatia. Novas estratégias terapêuticas dirigidas aos eventos intracelulares descritos deverão futuramente promover benefício adicional.
The determinant of the diabetic nephropathy is hyperglycemia, but hypertension and other genetic factors are also involved. Glomerulus is the focus of the injury, where mesangial cell proliferation and extracellular matrix occur because of the increase of the intra- and extracellular glucose concentration and overexpression of GLUT1. Sequentially, there are increases in the flow by the poliol pathway, oxidative stress, increased intracellular production of advanced glycation end products (AGEs), activation of the PKC pathway, increase of the activity of the hexosamine pathway, and activation of TGF-beta1. High glucose concentrations also increase angiotensin II (AII) levels. Therefore, glucose and AII exert similar effects in inducing extracellular matrix formation in the mesangial cells, using similar transductional signal, which increases TGF-beta1 levels. In this review we focus in the effect of glucose and AII in the mesangial cells in causing the events related to the genesis of diabetic nephropathy. The alterations in the signal pathways discussed in this review give support to the observational studies and clinical assays, where metabolic and antihypertensive controls obtained with angiotensin-converting inhibitors have shown important and additive effect in the prevention of the beginning and progression of diabetic nephropathy. New therapeutic strategies directed to the described intracellular events may give future additional benefits.
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Humans , Diabetic Nephropathies/etiology , Glomerular Mesangium , Hyperglycemia/complications , Angiotensin II/metabolism , Cell Proliferation/drug effects , Diabetic Nephropathies/metabolism , Diabetic Nephropathies/physiopathology , Endothelium-Dependent Relaxing Factors/metabolism , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Glomerular Mesangium/metabolism , Glomerular Mesangium/pathology , Glomerular Mesangium/physiopathology , Glucose Transporter Type 1/metabolism , /metabolism , Hyperglycemia/metabolism , Hyperglycemia/physiopathology , Nitric Oxide/metabolism , Oxidative Stress/drug effects , Renin-Angiotensin System/drug effects , Sclerosis/metabolism , Sclerosis/physiopathology , Transforming Growth Factor beta1/metabolism , Vasoconstrictor Agents/metabolismABSTRACT
Now the treatment of lymphoma mainly uses combined chemotherapy and radiotherapy.Es- pecially hemopoietic stem cell transplantation is effective in the treatment to the patients with refractory and later stage.During the recent years the domestic and foreign scholars have obtained the encouraging results with arsenic trioxide to treat lymphoma.This article has reviewed the experimental study and the clinical re- search progress about arsenic trioxide in application to lymphoma cells.