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1.
Article in Chinese | WPRIM | ID: wpr-744239

ABSTRACT

AIM:To investigate the role of SDF-1α/CXCR4 axis in pancreatic cancer cell migration and invasion.METHODS:The mRNA expression of CXCR4 in 4 pancreatic cancer cell lines was detected by RT-qPCR.The migration and invasion abilities of PANC-1 cells with the axis activated by exogenous SDF-1αor inhibited by CXCR4 inhibitor AMD3100 were detected by Transwell assays.The cell viability was measured by MTS assay.The protein expression of the epithelial-mesenchymal transition (EMT) -related molecules in the cells treated with exogenous SDF-1αor AMD3100 was determined by Western blot.RESULTS:All of the 4 pancreatic cancer cell lines expressed CXCR4 mRNA, while the PANC-1 cell line expressed the most.Exogenous SDF-1αpromoted the migration and invasion abilities of PANC-1 cells, which was inhibited by AMD3100.The PANC-1 cells treated with exogenous SDF-1αfor 72 h grew faster, while SDF-1αcombined with AMD3100 made little significance to the viability of PANC-1 cells.Exogenous SDF-1αinduced EMT of PANC-1 cells by up-regulating the expression of SNAIL and TWIST, and AMD3100 reversed this effect.CONCLUSION:SDF-1α/CXCR4 axis enhances the migration and invasion abilities of pancreatic cancer cells through inducing EMT.

2.
Article in Chinese | WPRIM | ID: wpr-699631

ABSTRACT

Objective To explore the role of integrin αvβ3 in the promotion of the development of choroidal neovascularization (CNV) by SDF-1/CXCR4.Methods This study was divided into two parts in vitro and in vivo.As for the in vivo study,a CNV model was induced by laser on C57BL/6J mice,and then assigned into 4 groups:mice with solely CNV modeling as control group,with intravitreal injection of SDF-1 after immediate CNV modeling as SDF-1 group,with intravitreal injection of SDF-1 + CXCR4 inhibitor (AMD3100) after CNV modeling as SDF-1 + AMD3100 group,and mice with intravitreal injection of SDF-1 + αvβ3 inhibitor (SB273005) after modeling as SDF-1 + SB273005 group.CXCR4 and αvβ3 expression levels in laser-induced eyes were quantified by qRT-PCR at time points of day 1,3,5,7,10 and 14 after modeling,and immunofluorescence staining was applied to detect αvβ3 expression in regional CNV and its endothelial cells in the four groups.Finally,OCT was used to observe the height of retinal pigment epithelial (RPE) layers in CNV after treatment in the four groups.Moreover,in the experiment in vitro,Western blot was used to measure the expression of CXCR4 protein of RF/6A cells in normal control group,Si-CXCR4 knockdown group and Si-NC knockdown model group.Meanwhile,the expression of integrin subunit β3 protein was determined in the normal control group,SDF-1 group,SDF-1 + AMD3100group,SDF-1 + Si-NC group and SDF-1 + Si-CXCR4 group.Transwell assay was conducted to detect the migration ability of RF/6A cells in the normal control group,SDF-1group,SDF-1 +AMD3100 group,SDF-1 + SB273005 group.Results On the one hand,the study in vivo,qRT-PCR showed that the expression of CXCR4 and integrin subunit β3 mRNA was up-regulated at first,and then down-regulated with time passed after CNV induction,with the highest expression level of CXCR4 mRNA (4.263 ± 0.464) on day 3,and the peak expression of β3 mRNA (3.678 ±0.364) on day 7 after CNV modeling.The results of immunofluorescence staining showed that the β3 fluorescence intensity of SDF-1 group was significantly enhanced,and the ratio of β3/CD31 was also significantly increased,which both were significantly higher than those of the control group (P < 0.01).However,the β3 fluorescence intensity and β3/CD31 ratio of SDF-1 +AMD3100 group and SDF-1 + SB273005 group were significantly weakened and decreased,respectively (P <0.05).OCT showed that the elevation level of RPE layer inSDF-1 group was significantly higher than that in the control group [(135.503 ± 10.301) μm vs.(94.443 ± 12.156) μm](P<0.05).The height of RPE uplift in SDF-1 + AMD3100 group [(95.283 ±20.062) μm] and SDF-1 + SB273005 group [(99.807 ± 10.403) μm] was significantly decreased (P < 0.05).On the other hand,in experiment in vitro,Western blot showed that the expression levels of integrin β3 in SDF-1 group and SDF-1 + Si-NC group were significantly higher than those in the control group [(1.301 ± 0.043) and (1.273 ± 0.077) vs.(0.244 ± 0.069)] (P < 0.01).The levels of integrin subunit β3 protein in SDF-1 + si-CXCR4 group (0.322 ± 0.042) and SDF-1 + AMD3100 group (0.336 ± 0.077) were significantly down-regulated (P < 0.01).Transwell assay showed that the amount of migrating cells in SDF-1 group increased,which was significantly higher than that of the control group (P < 0.01),while the number of migrating cells in SDF-1 +AMD3100 group and SDF-1 + SB273005 group was significantly decreased.Conclusion Integrin αvβ3 can promote the development of CNV by mediating SDF-1/CXCR4 signaling in endothelial cells.

3.
Article in Chinese | WPRIM | ID: wpr-620026

ABSTRACT

Objective To investigate the effects of stromal cell-derived factor 1 (SDF-1) and an CXC chemokine receptor 4 (CXCR4) antagonist (AMD3100) on the invasion and migration capabilities of the huaman choriocarcinoma cell line JAR for further elucidating the role of SDF-1/CXCR4 axis in the pathogenesis of preeclampsia.Methods JAR cells were divided into four groups: SDF-1 group (treated with 50 ng/ml of SDF-1),SDF-1+AM3100 mixed group (first treated with 100 ng/ml of AMD3100 for 2 hours and then treated with 50 ng/ml of SDF-1),AMD3100 group (treated with 100 ng/ml of AMD3100) and blank control group (without any treatment).RT-PCR was performed to detect the expression of CXCR4 at mRNA level in JAR cells.Western blot assay was used to measure the expression of CXCR4 and p-AKT at protein level.MTT assay was used to analyze the effects of different concentrations of SDF-1 (10,30,50 and 100 ng/ml) on the proliferation of JAR cells at different time points (0,24,48,72 h).Transwell invasion assay and wound-healing assay were used to test the changes in invasion and migration capabilities of JAR cells after different treatments.Results (1) Results of the RT-PCR showed that the expression of CXCR4 at mRNA level in JAR cells was increased in the SDF-1 group (1.839±0.083) as compared with that in the blank control group (1.372±0.086),AMD3100 group (0.694±0.045) or SDF-1+AM3100 mixed group (0.703±0.093).Moreover,the differences between the SDF-1 group and the other three groups were statistically significant (F=30.67,P<0.05).Compared with the blank control group,the expression of CXCR4 at mRNA level in JAR cells was decreased in the AMD3100 group (P<0.01).(2) Results of the Western blot assay showed that the expression of CXCR4 and p-AKT at protein level in JAR cells were enhanced in the SDF-1 group as compared with that in the blank control group,AMD3100 group or SDF-1+AM3100 mixed group.Compared with the blank control group,the expression of CXCR4 and p-AKT at protein level in JAR cells were inhibited in the AMD3100 group.(3) Results of the MTT assay showed that SDF-1,especially at the concentration of 50 ng/ml,could enhance the proliferation of JAR cells (P<0.05) and its best effect on proliferation was seen at 48 h.(4) Results of the Transwell invasion assay showed that the number of transmembrane cells in the SDF-1 group (70.49±2.42) was more than that in the blank control group (54.36±2.26),AMD3100 group (21.68±8.31),or SDF-1+AMD3100 mixed group (28.18±4.61).The differences between the SDF-1 group and the other three groups were statistically significant (F=116.26,P<0.01).Compared with the blank control group,the number of transmembrane cells was reduced in the AMD3100 group (P<0.05).(5) Results of the wound-healing assay showed that the relative migration distance was increased in the SDF-1 group (1.162±0.034) as compared with that in the blank control group (0.823±0.101),AMD3100 group (0.160±0.047),or SDF-1+AMD3100 mixed group (0.183±0.064).The differences between the SDF-1 group and the other three groups were statistically significant (F=30.500,P<0.05).Compared with the blank control group,the relative migration distance was decreased in the AMD3100 group (P<0.01).Conclusion The invasion and migration of huaman choriocarcinoma JAR cells can be enhanced by SDF-1,but inhibited by AMD3100.This study indicates that the blocked biological axis of SDF-1/CXCR4 may play an important role in the pathogenesis of preeclampsia through inducing abnormal activation of PI3K/AKT pathway,which results in inhibited invasion and migration of trophoblast cells and placenta abnormality.

4.
Basic & Clinical Medicine ; (12): 1529-1534, 2017.
Article in Chinese | WPRIM | ID: wpr-666989

ABSTRACT

Objective To investigate the role of SDF-1/CXCR4 axis on the apoptosis of human degenerative nucleus pulposus cells (NPCs) and its potential molecular mechanism .Methods The intervertebral disces tissues from clinical discectomy were divided into normal group and intervertebral disc degeneration ( IVD) group according to Pfirrmann classification.The different expression of SDF 1 and CXCR4 in human IVDs was tested by immunohistochemistry, quantify polymerase chain reaction (q-PCR) and Western blot.The primary degenerative NPCs were primary cultured.The generation Ⅲ~Ⅴ NPCs was treated with 10 ng/mL SDF-1, in the presence of or in the absence of CXCR4 siRNA transfection and 20 μmol/L NF-κB inhibitor (pyrrolidine dithiocar bamate,PDTC).The transfection efficiency and target protein of signal pathway were verified by Western blot , the apoptosis of NPCs were tested by Annexin V /PI, the nucleus transferlocation of P65 from NF-κB were tested by immunofluorescent method.Results SDF-1and CXCR4 were both expressed in all donor tissues, however, there was a significantly increased in the degenerative IVDs .The apoptosis of degenerative NPCs was expedited by SDF -1 stimulation,which was significantly suppressed by CXCR 4 silencing by siRNA (P<0.05).Furthermore, with SDF-1 stimulation,the expressions of phosphorylated P 65 was significantly increased and the P65 perssad transferred to the nucleuses,which could be suppressed by the NF-κB inhibitor, PDTC(P<0.05).Conclusions The expression levels of SDF-1 and CXCR4 are increased in degenerative NP tissue.The SDF-1/CXCR4 axis is considered to induce apoptotic of human degenerative NPCs via the NF-κB signaling pathway.

5.
Chinese Pharmacological Bulletin ; (12): 427-432,433, 2016.
Article in Chinese | WPRIM | ID: wpr-603570

ABSTRACT

Aim To investigate the role of Xuefuzhuyu decoction ( XFZYD ) combined with EPC in repairing damaged vascular endothelium using traditional Chi-nese medicine way of blood circulation combined with cell therapy. Methods The repaired situation of inju-ried endothelium was observed and the effect of XFZYD on EPC was analysed after the endothelial in-juried rats were gavaged XFZYD and vena caudalis in-jected EPC. Results Compared with EPC group and XFZYD group, the XFZYD joint EPC group ’ s endo-thelial thickness was reduced significantly(P<0. 05). And there appeared more significant role in lowering triglycerides, total cholesterol and increasing HDL lev-els( P<0. 05 ) , the calcium was decreased more sig-nificantly( P <0. 05 ); vascular eNOS protein expres-sion increased significantly(P<0. 05); vascular SDF-1 expression was significantly increased. Conclusion XFZYD can promote EPC repairing damaged endotheli-um, and the mechanism may be relevant to improving the environment and promoting the EPC homing.

6.
Tianjin Medical Journal ; (12): 457-460, 2015.
Article in Chinese | WPRIM | ID: wpr-473832

ABSTRACT

Objective To investigate the molecular regulatory mechanism of glucagon like peptide 1 (GLP-1) on proliferation, differentiation and apoptosis of human umbilical cord blood endothelial progenitor cells (EPCs). Methods EPCs were isolated from the umbilical cord blood of healthy pregnant women and cultured in 6-hole cell plate at 2×105 density in vitro, transfected with empty vector plasmid (control group), pcDNA3-GLP-1 plasmid (GLP-1 group), pcDNA3-GLP-1plasmid+AMD3100 (GLP-1+AMD3100 group) and simple AMD3100 (AMD3100 group). The pcDNA3-GLP-1 was transfected into EPCs. The 25μmol/L AMD3100 was used to block the SDF-1/CXCR4 signal pathway of EPCs for 1 h. The cell proliferation was determined by MTT method. The mRNA expressions of differentiation and apoptosis related genes PPARγ, C/EBPα and Caspase-3 were investigated by RT-PCR, and Caspase-3 activity was determined by Caspase-3 activity assay kit. Results Compared to control group, AMD3100 inhibitor showed no effects on cell proliferation, differentiation and apoptosis, while over-expression of GLP-1 in EPCs obviously promoted cell proliferation, and differentiation related genes PPARγand C/EBPαmRNA expression, but down-regulated mRNA expression and the activity of Caspase-3 significantly (P<0.05), indicating that GLP-1 increased proliferation and differentiation of EPCs while decreased cell apoptosis. When the SDF-1/CXCR4 signaling pathway was blocked by AMD3100, over-expression of GLP-1 induced promotion of cell proliferation, and the differentiation was decreased significantly and the apoptosis was significantly increased (P<0.05). Conclusion These data confirm that GLP-1 might promote EPCs proliferation and differentiation, and inhibit cell apoptosis through the regulation of the SDF-1/CXCR4 signaling pathway.

7.
Article in Chinese | WPRIM | ID: wpr-576082

ABSTRACT

Objective:To investigate the effects of anti-CXCR4 monoclonal antibody 12G5 on adhesion and invasion of SGC7901 cells.Methods:The effects of 12G5(5?g/ml,10?g/ml and 20?g/ml, respectively)on the adhesive、invasive and migrative abilities of SGC7901 cells were explored by MTT and Transwell chamber. Results:(1)12G5 showed a time/dose-dependent inhibition effect on SGC7901 cells adhering Matrigel,and the adhesion of gastric cancer cells was inhibited on the concentration of 10?g/ml of 12G5, and compared with untreated group,adhesion of SGC7901 cells was signficantly lower(P

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