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Objective The aim of this study was to observe the expression of human epidermal growth factor receptor( EG-FR),epidermal growth factor receptor 2(HER2)and epidermal growth factor receptor 3(HER3)in cholangiocarcinoma(CCA),and to explore the relationship between the expression of EGFR,HER2,HER3 and the clinicopathological features,overall survival time and proliferation of CCA patients. Methods Fifty patients with CCA who underwent surgery in our hospital from January 2009 to October 2013 were enrolled,and their adjacent tissues were also collected as controls. Immunohistochemistry and qRT-PCR were used to de-tect the expression of EGFR,HER2 and HER3 in CCA and adjacent tissues. The clinicopathological parameters of patients were col-lected and the relationship between EGFR,HER2,and HER3 expression and clinicopathological parameters of CCA was analyzed. Ka-plan-Meier survival curves were plotted,and the relationship between the expression of EGFR,HER2,and HER3 and total postopera-tive survival of 50 patients with CCA was analyzed using the Log-rank test and Cox proportional hazard model. The expression of EG-FR,HER2 and HER3 in CCA QBC939 cell line was knocked down by RNA interference assay. The knockdown effect of EGFR,HER2 and HER3 was detected by Western blot. The effect of EGFR,HER2 and HER3 on proliferation of QBC939 cells was determined by MTT assay. Results The positive expression of EGFR,HER2 and HER3 was determined in CCA tissues. The relationship analysis of clinicopathological characteristics showed that the HER2 expression was associated with CCA lymph node metastasis(P<0. 05). EG-FR and HER3 was associated with CCA lymph node metastasis and correlated with cancer differentiation(P<0. 05). The overall sur-vival of patients with EGFR,HER2 and HER3 positive was significantly lower than that of negative patients( P<0. 05). After knoc-king EGFR,HER2 and HER3 expression,the proliferation was significantly decreased in QBC939 cells(P<0. 05). Conclusion The expression of EGFR,HER2 and HER3 in CCA tissues is closely related to the overall survival of patients,and the mechanism may be related to the promotion of proliferation of CCA cells.
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Objective To explore the clinical value of serum vascular endothelial grow th factor(VEGF), γ-glutamyl transferase(GGT)and soluble myeloid cell trigger receptor-1(sTREM-1)in early chronic obstruc-tive pulmonary disease(COPD).Methods 30 cases of COPD patients admitted from February 2013 to Febru-ary 2015 were selected as the observation group,and 30 healthy subjects were selected as the control group. The levels of serum VEGF,GGT and sTREM-1 were compared between the two groups.Results The levels of serum VEGF,GGT and sTREM-1 were significantly decreased in the observation group after treatment,the difference was statistically significant(P<0.05).The levels of serum VEGF,GGT and sTREM-1 in the obser-vation group were significantly higher than those in the control group(P<0.05).The serum VEGF,GGT and sTREM-1 were respectively detected under the ROC curve area was 0.83,0.80,0.67,specificity and sensitivi-ty were 73.3%,71.1%,58.9% and 91.3%,88.6%,80.2%.ROC curve area of combined detection of VEGF, and GGT in early diagnosis of COPD was 0.94,the sensitivity and specificity were 81.3% and 96.4%;ROC curve area of combined detection of VEGF,and sTREM-1 in early diagnosis of COPD was 0.89,the sensitivity and specificity were 75.5% and 92.8%.Conclusion Serum VEGF,GGT,sTREM-1 level can be used as an important monitoring index for early diagnosis of COPD.
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Objective To study the regulation of microRNA - 22(miR - 22)on glycometabolism of hemato-poietic stem cell TF - 1 and its molecular mechanism. Methods TF - 1 cells were cultured for 2 h under hypoxic con-ditions. The expression levels of Glut4 and miR - 22 was detected by quantitative real-time PCR(qRT - PCR). The sgRNA vector of the miR - 22 gene was constructed and miR - 22 gene of TK - 1 cells was knocked out by the CRISPR/ Cas9 technique. Overexpression vectors were constructed,and miR - 22 knocked - out cells were introduced to overexpress miR - 22,the expression of miR - 22 was detected by qRT - PCR and the expression levels of Glut4 and PPAR - γ were detected by qRT - PCR and Western blot. Results Compared with the control group,the expression of miR - 22 in TF - 1 cells decreased (0. 015 ± 0. 000 vs. 0. 056 ± 0. 001)and the expression of Glut4 (0. 351 ± 0. 038 vs. 0. 152 ± 0. 005)and PPAR - γ (0. 421 ± 0. 017 vs. 0. 248 ± 0. 008)increased,when TF - 1 cells were cultured for 2 h under hypoxic conditions,and the differences were statistically significant (all P < 0. 05). Compared with the control group,the expression levels of Glut4 (0. 019 ± 0. 00 vs. 0. 008 ± 0. 000)and PPAR - γ (0. 038 ± 0. 001 vs. 0. 019 ± 0. 000)were significantly increased after miR - 22 gene silencing,and they were significantly decreased (Glut4:0. 005 ± 0. 000 vs. 0. 008 ± 0. 000;PPAR - γ:0. 137 ± 0. 000 vs. 0. 019 ± 0. 000)after overexpression of miR - 22,and the differences were statistically significant (all P < 0. 05). Conclusions It suggests that miR - 22 ex-erts a negative regulation on glycometabolism of hematopoietic stem cell TF - 1 by downregulating the expression of PPAR - γ. A new regulatory factor of TF - 1 glycometabolism and the mechanisms are identified,which has provided new ideas for the targeted medication of diseases induced by hematopoietic stem cell dysfunction.
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Objective To investigate the effect of bevacizumab combined with chemotherapy on the expression of EGFR and HER in the patients with metastatic colorectal cancer .Methods Sixty patients with metastatic colorectal cancer treated in this hos-pital from January 2013 to October 2016 were selected and randomly divided into the experiment group (bevacizumab combined with FOLFOX-6 chemotherapy regimen) and control group (FOLFOX-6 chemotherapy regimen) 30 cases in each group .The curative effects were observed in the two groups .The levels of serum HER and EGFR were detected by enzyme-linked immunosorbent assay (ELISA) .Results The total effective rate after treatment in the experiment group was 53 .33% ,which in the control group was 33 .33% ,the difference was statistically significant (P< 0 .05) .After treatment ,the levels of serum HER and EGHR in the two groups were decreased to some extent ,moreover the experiment group had more decrease than the control group ,the difference be-tween the two groups was statistically significant (P<0 .05) .The occurrence rate of adverse reactions had no statistical difference between the two groups (P>0 .05) .Conclusion Bevacizumab combined with chemotherapy has good effect in the patients with metastatic colorectal cancer .
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[Objective] To observe the effect of Jiedu Quyu Ziyin decoction on TLR4 signaling pathway in macrophages of MRL/lpr lupus mice. [Method] MRL/lpr lupus mice were divided into four groups:model group, prednisone group, Jiedu Quyu Ziyin decoction group(hereinafter referred to as:Chinese medicine group) and prednisone plus Chinese medicine group (hereinafter referred to as:combination of Chinese and western medicine group). The mice were gavaged with saline, prednisone, Jiedu Quyu Ziyin decoction and prednisone added Jiedu Quyu Ziyin decoction for 4 weeks. Macrophages of lung, peritoneal and spleen were collected and the expression of related genes was detected by RT-PCR. [Result] TLR4 mRNA in lung macrophages, and TLR4 protein in splenic macrophages increased significantly(P<0.05) after the treatment of prednisone. The increased TLR4 protein in splenic macrophages was significantly decreased by combining with Jiedu Quyu Ziyin decoction(P<0.05). Prednisone can significantly reduce the TLR4 downstream molecules such as MyD88, IFN-α, iNOS mRNA expression in lung and peritoneal macrophages(P<0.05). The decreased MyD88 mRNA in lung macrophages was increased significantly by combining with Jiedu Quyu Ziyin decoction(P<0.05). [Conclusion] TLR4 signaling pathway is changed in macrophages after glucocorticoid administration in MRL/lpr lupus mice. Jiedu Quyu Ziyin decoction can reduce the abnormal glucocorticoid-induced TLR4 protein expression.
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AIM:To investigate the role of canonical transient receptor potential channel 1 ( TRPC1 ) in the migration of human bronchial epithelial cells (16HBE) induced by transforming growth factor-β1 ( TGF-β1).METH-ODS:Silencing of TRPC1 gene expression was performed by siRNA.The cell activity and apoptosis were measured by CCK-8 assay and flow cytometry, respectively.The migration and invasion abilities of the 16HBE cells were detected by wound-healing assay and Transwell assay.The protein expression of E-cadherin and vimentin was determined by Western blot.RESULTS:TGF-β1 treatment significantly enhanced the cell migration distance compared with control groups ( P0.05).The results of wound-healing and Tr-answell assays showed that migration and invasion abilities in TRPC1 siRNA +TGF-β1 group were markedly suppressed compared with TGF-β1 group (P<0.01).The protein expression of E-cadherin and vimentin induced by TGF-β1 was in-hibited by TRPC1 silencing compared with TGF-β1 group (P<0.05).CONCLUSION:TRPC1 is involved in the migra-tion of human bronchial epithelial cells induced by TGF-β1 through regulating the protein expression of E-cadherin and vim-entin.
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Breast cancer has become the most common malignant tumor and the major cause of cancer-related death for women around the world. The number of patients shows an increasing trend recently. Breast cancer is a big threaten to wom?en’s health and quality of life. With the development of molecular biology, molecular biomarkers have been found assiciated with prognosis in patients with breast cancer, which makes it possible to predict cancer patient survival precisely and practi?cally. This review summarized those new developments of biomarkers on the prognosis of breast cancer.
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Aim To investigate the selective inhibition of ethanol on muscarinic receptor-or 5-HT receptor-me-diated contractile responses in the circular smooth mus-cle strips isolated from the different regions of rat stom-ach. Methods Circular muscle strips isolated from the rat gastric fundus, body, cardia and pylorus were prepared, and the contractile responses to carbachol ( CCh ) or 5-HT were recorded. Results Ethanol (0. 000 05~0. 000 5, V/V) did not affect the contrac-tile response to CCh in circular muscle strips from the rat gastric fundus and cardia, and that to 5-HT in the strips from rat gastric fundus and body ( P >0. 05 ) . However, ethanol(0. 000 1 and 0. 000 5) significantly inhibited the Emax value of the contraction by CCh from (12. 18 ± 0. 33) g of control level to (10. 88 ± 0. 41) g and -lgEC50 value from ( 6. 33 ± 0. 05 ) of control level to (6. 12 ± 0. 06)(P <0. 05) in the strips from rat gastric body. Ethanol(0. 000 1 and 0. 000 5) also significantly inhibited the Emax value of the contraction by CCh from (2. 87 ± 0. 15) g of control level to (2. 2 ± 0. 13) g and -lgEC50 value from (6. 49 ± 0. 10) of control level to (6. 05 ± 0. 09)(P<0. 01) in the strips from rat gastric pylorus. Moreover, ethanol ( 0. 000 1 and 0. 000 5) significantly inhibited the Emax value of the contraction by 5-HT from (2. 93 ± 0. 35) g of con-trol level to ( 2. 1 ± 0. 30 ) g ( P<0. 05 ) , but did not affect the -lgEC50 value in the strips from rat gastric cardia. Conclusions Ethanol inhibits the contractile responses to 5-HT only in the circular muscle strips of rat gastric cardia, and it inhibits the contractile respon-ses to CCh more strongly in the circular muscle strips of gastric pylorus than gastric body. In those gastric circular muscle strips, ethanol decreases both the ac-tivity and affinity of CCh to muscarinic receptors, but decreases only the activity of 5-HT to its receptors.
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Objective To study the effect of brain‐derived neurotrophic factor (BDNF) in the process of adenosine triphos‐phate (ATP) activating BV2 microglia .Methods BV2 microglia was cultured by adding different concentrations of ATP .Then the expression level of intracellular CD11b and BDNF and the secretion level of TNF‐α in the supernatant were quantitatively deter‐mined by Western blot .BV2 microglia was treated by different concentrations of BDNF scavenger tyrosine kinase receptors B (TrkB)/Fc and incubated by ATP .The expression level of intracellular CD11b and BDNF and the secretion level of TNF‐αin the supernatant were measured .Finally adding exogenous recombinant BDNF into cultured BV 2 microglia ,intracellular changes of CD11b and supernatant TNF‐αlevels were detected .Results After adding ATP for cultivating BV2 microglia ,intracellular CD11b and BDNF expression levels and supernatant TNF‐αlevel were increased with a dose‐and time‐dependent manner in some ranges . After adding TrkB/Fc ,the levels of intracellular CD11b and BDNF expression and supernatant TNF‐αlevel were decreased with a dose‐and time‐dependent manner in some range .CD11b and BDNF expression levels was decreased in a dose‐and time‐dependent manner .Adding exogenous BDNF ,the levels of intracellular CD11b and BDNF expression and supernatant TNF‐α level were in‐creased again .Conclusion Intracellular BDNF expression is increased when BV2 microglia is activated and replenishing exogenous BDNF can activate microglia .Therefore BDNF may play an important role in the microglia activation process .
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To clarify the molecular mechanism of SAHA in the cell proliferation of ER-positive breast cancer cell line MCF-7 induced by leptin. Methods Human breast cancer cell MCF-7 was incubated with SAHA and/or leptin, and cell viability, apoptosis and cell cy-cle of MCF-7 cells were detected by Muse Cell Analy-zer. The expression of proteins related with apoptosis was determined by apotosis antibody array. Results Real-time cell proliferation assays indicated that the in-duction effect of leptin for MCF-7 cells reached the peak at a concentration of 0. 625 nmol · L-1 . SAHA reduced the viability of MCF-7 cells, induced G0/G1 phase arrest in the cell cycle, and triggered the apopto-sis. Meanwhile, SAHA significantly induced the pro-tein expressions of some apoptotic factors, including Bax, Caspase-3, TRAIL DR5, p21CIP1, and inhibited the expressions of Claspin, Clusterin, x-linked inhibi-tor of apoptosis protein(XIAP) and survivin. Howev-er, leptin had reverse effects on the related expression of the proteins. Conclusion The effects of cell prolif-eration by leptin and SAHA treatment in breast cancer ER positive cell line MCF-7 may involve in the activa-tion of apoptosis pathway, in particular with releasing of Caspase-3 trigged by endogenous mitochondrial ap-optosis pathway.
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The fibrosis can occur in many kinds of organs,and its sustained progress may lead to organ structural damage and functional decline,and even the organ failure,which threatens the human health and the life seriously.Adenosine is an endogenous purine nucleoside that can be generated in various tissues of the body and regulate a multitude of body functions via the combina-tion with four different kinds of G protein-coupled receptors.Re-cent studies have found that adenosine receptors play an impor-tant role in regeneration tissue and fibrosis process.To under-stand the processes may be helpful to the treatment of fibrosis diseases.This review makes a summary on latest research pro-gress of adenosine receptors in fibrosis diseases.
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Aim To investigate the estrogenic activities of four components from pregnant mare’s urine extract. Methods The estrogenic activities of four components were assessed using two in vitro tests:the MCF-7 cell proliferation assay (E-screen test)and the luciferase transfected CHO cell gene reporter assay.In the lucifer-ase reporter gene assay,the reporter gene plasmids PGM-ERE-Luc and ERαor ERβand a control plasmid (pRL-cmv)were transiently co-transfected into CHO cells to establish an ERα-or ERβ-cell screening system which was used to measure estrogenic activity of four compounds.Results MCF-7 cells treated with HP, DHP,PT and HA significantly proliferated,thereby of-fering in vitro evidence for the estrogenic activities of HP,DHP,PT and HA,and they showed dose-depend-ent activities.Compared EC50 of PE and RPE with that of E2 ,HP,DHP,PT and HA exerted relatively weak estrogenic activities.The in vitro ER-mediated reporter gene assay revealed that HP,DHP,PT and HA dis-played estrogenic activities mediated by ERβor ERα. Compared with the EC50 of E2 ,HP,DHP,PT and HA exhibited lower estrogenic potencies.Conclusion HP, DHP,PT and HA possess weaker estrogenic activities than E2 .
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Aim To explore the effects of tanshinone IIA on cell proliferation via G protein-coupled estrogen receptor inductive and regulative pathway in typical es-trogen receptor and G protein-coupled estrogen receptor positive T47D breast cancer cells. Methods The pro-liferation rate of T47 D cells influenced by tanshinone IIA was analyzed by MTT assay. G protein-coupled es-trogen receptor agonist G1 and GPER antagonist G15 were employed as tools. GPER SiRNA was applied to build GPER gene silence T47D cells. GPER expres-sion influenced by tanshinone IIA was measured by Western blot. Results The proliferation rates of T47D cells treated with 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol· L-1 of tanshinone IIA were decreased significantly. Such effects could be attenuated by G1 or enhanced by G15 . Growth of GPER SiRNA transfected T47 D cells were significantly inhibited by 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol·L-1 of tanshinone IIA treating. Result of Western blot showed that tanshinone IIA at 1 × 10 -5 mol· L-1 and 1 × 10 -6 mol · L-1 could induce de-crease of GPER protein expression in T47D cells. Conclusions Tanshinone IIA shows inhibitory effects on proliferation rate of T47 D breast cancer cells via GPER pathway. Tanshinone IIA could perform regula-tive function on GPER expression level in target cells.
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Sphingosine-1-phosphate receptor 1(S1PR1) is a new member of G protein-coupled receptor family. A great body of data suggest S1PR1 is capable of regulating lots of downstream signaling molecules and cellular processes. It is found that S1P/S1PR1 plays an important role in the development and mainte-nance of pain. However, it is controversial whether activation of S1PR1 would enhance or attenuate pain. Here, recent studies ＜br> and current perspectives are discussed in order to better under-stand the biological and pathological roles of S1PR1 in pain mod-ulation.
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Objective To analyze the differences with the expression of glucocorticoids receptor isoforms ( GRα, GRβ, GRγand GR-P ) and cytokines [ IL-6, macrophage migration inhibitory factor (MIF), IFN-γand IL-10] between patients with systemic lupus erythematosus (SLE) and rheumatoid ar-thritis ( RA) , and to further understand their correlations with disease activities.Methods Fifty-five pa-tients with SLE, forty-nine patients with RA and thirty-eight healthy subjects were recruited in this study. All patients were steroid-naive.The expression of GRα, GRβ, GRγ, and GR-P in peripheral blood mononu-clear cells at transcript levels were determined by real-time PCR.Enzyme-linked immunosorbent assay was used to detect the expression of IL-6, MIF, IFN-γand IL-10 in serum samples.Results The percentages of GRαin all subjects were the highest among four isoforms of GR, followed by GR-P, GRγand GRβ.Com-pared with healthy subjects, patients with SLE or RA showed significantly decreased expression of GRα( P<0.05), but increased expression of GR-P (P<0.05).The percentages of GR-P in patients with RA were higher than those in patients with SLE (P<0.05).The expression of GRαwas negatively correlated with SLE disease activity index (SLEDAI) and disease activity score 28 (DAS28).SLE or RA patients with high disease activity showed lower expression of GRαthan those with low disease activity.The levels of IL-6, IFN-γand MIF in patients with SLE or RA were significantly higher than those in healthy subjects ( P<0.05).A negative correlation was observed between the expression of IL-6 and GRαin patients with SLE (P<0.05).The expression of IFN-γwas negatively correlated with GRαin patients with RA (P<0.05). Conclusion There were significant differences with the expression of GR isoforms among patients with SLE, patients with RA and healthy subjects, indicating the change of internal environment in patients might be in-volved in GR alternative splicing.GRαwas the predominant isoform and was negatively correlated with dis-ease activities.Oversecretion of cytokines resulted in a decreased expression of GRα.This study would be useful for the diagnosis of the disease status and for monitoring clinical treatment.
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Aim Toinvestigatewhetherepidermal growth factor receptor ( EGFR )-containing exosomes could induce tumor-specific regulatory T cells, and the effects of those T cells on tumor protein-specific CD8+Tcells.Methods TheexosomeswithEGFRwerepu-rified from NSCLC tumor, which modulated tolerogenic property of DCs. Then the induced TolDCs generated tumor-specific Tregs, with which the tumor protein-specific CD8 + T cells were suppressed. Results 80%exosomeswereEGFRpositivefromLCpatients while less than 2% exosomes were EGFR positive from control lung tissue. After exposed to the exosomes in the culture for 7 days, the IDO+ DCs proportion was much higher than that in control group ( 80. 8% ± 3. 2% vs 65. 6% ± 6. 4%, P <0. 05 ) . The induced Tregs was also higher ( 24. 1% ± 5. 2% vs 4. 2% ± 2. 3%,P<0. 01 ) , which suppressed the proliferation of CD8+ T cells(5. 4% ± 0. 2% vs 86. 7% ± 9. 3%, P<0.01).Conclusion Thepurifiedexosomesin-duce tolerogenic DCs. Coculture of the tolerogenic DCs and Th0 cells generates the tumor antigen-specific reg-ulatory T cells. The Tregs could suppress the tumor an-tigen specific CD8+ T cells.
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AngiotensinⅡ( AngⅡ) is the main functional bioac-tive peptide of the renin angiotensin system,and its basic function is to regulate salt-water homeostasis and blood pressure. Howev-er,recent studies have shown that AngⅡ plays a very important role in tumor formation and angiogenesis by acting on its type 1 receptor (AT1R) as a kind of potential growth factor. This arti-cle is a brief overview of the research on effects of AngⅡ-AT1R system in the process of tumor angiogenesis in recent years.
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G-protein coupled receptor (GPCR) family is the large st group of therapeutic targets. High throughput screening (HTS) plays critical ro les in early stage of drug discovery. Based on signaling pathway stimulated by l igand binding on the GPCRs, many functional assay technologies have been develop ed for HTS. These technologies include Fluorometric microvolume assay technology (FMAT), Fluorescence polarization (FP), Competitive enzyme-linked immunosorben t assay (ELISA), Scintillation proximity assay (SPA), Melanophore assay, Reporte r gene assay and Calcium assay. The principles and applic- ations of these technologies were summarized in this review. We particularly emphasize those techniques of nonradioactive, su bstrate and cofactor free assays, which will be the major approaches for HTS, su ch as reporter gene and calcium assays.
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Aim To visualize the dynamics of PtdIns(4,5)P 2 hyd ro lysis and resynthesis, and modulate it by pharmacological agents wortmannin, LiC l, U73122 and neomycin. Methods We used a fusion construct of g reen fluorescent protein(GFP) with the PH domain of phospholipase C ?1(PL C ?1PH)(PLC ?1PH-GFP) known to bind PtdIns(4,5)P 2 specifically , and laser-scanning confocal microscopy to trace PtdIns(4,5)P 2 translocatio n. Results Stimulation of endogenous P 2Y receptors by ATP in CHO cells induced a reversible PLC ?1PH-GFP translocation, indicating Pt dIns(4,5)P 2 hydrolysis through the receptor-mediated phospholipase C (PLC) ac tivation. Wortmannin and LiCl did not affect the translocation of PLC ?1PH -GFP from plasma membrane to cytosol but blocked the recovery after the translo cation. The transient translocation from plasma membrane was blocked by the PLC inhibitor U73122 but was not affected by another PLC inhibitor neomycin. However , in the absence of PLC ?1PH-GFP expression, neomycin inlibited the recep tor-induced PLC hydrolysis of PtdIns(4,5)P 2.Conclusion PLC ?1PH-GFP can be used as a valuable fluorescence probe to visualize the dyn amic change of PtdIns(4,5)P 2 in living cells. Wortmannin, LiCl, U73122 and neo mycin have distinct modulation effects on PtdIns(4,5)P 2 metabolism. PLC ?1 PH,when bound to PtdIns(4,5)P 2,prevents neomycin from inhibiting PLC hydro lyzing PtdIns(4,5)P 2.