1.Effects of interferon-γ on expression of adhesion molecules in human umbilical cord mesenchymal stromal cells.
Rui-Yun QIAO ; Hai BAI ; Cun-Bang WANG ; Jian-Feng OU ; Hai-Ying ZHANG ; Qiang ZHAO
Journal of Experimental Hematology 2012;20(5):1195-1199
This study was purposed to investigate the effects of interferon (IFN)-γ on expression of adhesion molecules in mesenchymal stromal cells derived from human umbilical cord tissue (UC-MSC). The UC-MSC were isolated from human umbilical cord by tissue culture. The expressions of specific markers on UC-MSC were detected by flow cytometry in the physiological condition. The adipogenic and osteogenic induction of UC-MSC was detected by alizarin and Oil red O staining. UC-MSC were exposed to IFN-γ (100, 1 000, 10 000 U/ml) for 24 h, the expressions of CD54, CD58, CD44, CD49d, CD62p, CD62L, CD102 and CD106 on cell surface were detected using flow cytometry. The results showed that in physiological condition, UC-MSC extremely low expressed CD102, CD106, CD62P, CD62L, while the expression of CD54 was relatively high (41.58 ± 0.83)%. When stimulated by IFN-γ, the expression of CD102, CD106, CD62P, CD62L increased slightly, but still low (< 5%), while CD54 and CD58 upregulated concentration-dependently up to (59.66 ± 1.36)% and (43.96 ± 0.62)% respectively. The expression of CD49d upregulated to (51.33 ± 0.74)% when UC-MSC exposed to IFN-γ 100 U/ml. CD44 increased to (73.22 ± 1.93)% when UC-MSC exposed to IFN-γ 1 000 U/ml. It is concluded that IFN-γ can elevate significantly the expression of CD54, CD49d, CD44 and CD58, but has no significant effect on CD102, CD106, CD62P and CD62L expression on the surface of UC-MSC.
Cell Adhesion Molecules
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metabolism
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Cells, Cultured
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Humans
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Interferon-gamma
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pharmacology
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Mesenchymal Stromal Cells
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cytology
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drug effects
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metabolism
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Umbilical Cord
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cytology
2.Effect of platelet factor 4 on the adhesive property of leukemic CD34+ KG1a cell.
Jing ZHANG ; Yue-xia MA ; Zhong-chao HAN
Acta Academiae Medicinae Sinicae 2002;24(2):160-164
OBJECTIVETo study the effect of PF4 on the adherence of leukemic CD34+ KG1a cell to human umbilical vein endothelial cell line ECV-304 cell and on the expression of adhesive molecules.
METHODSAdhesion assay and adhesion blocking assay were respectively applied to measure the effect of PF4 and/or adhesion molecule monoclonal antibodies on the adhesion property of KG1a. The expressions of adhesion molecules were determined by RT-PCR and FACS analysis.
RESULTSThe adhesion of KG1a cells to ECV-304 was significantly enhanced in the presence of PF4. Such enhancement was also observed when KG1a or ECV-304 cells were separately treated with PF4 before interaction. The adhesion capacity of KG1a cells was reduced when cells were co-incubated with the blocking monoclonal antibodies (MoAbs) against CD49d, CD106, CD54, respectively. In contrast, MoAbs against CD62L, CD62P and CD62E had no such effect. During a period of 3 hours when KG1a or ECV-304 cells were respectively incubated with PF4, the mRNA expressions of CD49 d, CD54 were up-regulated. Furthermore, when KG1a or ECV-304 cells were incubated with PF4 for 2 hours, respectively, the percentages of CD49d+ KG1a cells and CD54+ ECV-304 were increased significantly.
CONCLUSIONPF4 can enhance KG1a cell adhesive capacity by increasing the expressions of adhesion molecules.
Antigens, CD34 ; metabolism ; Cell Adhesion ; drug effects ; Cell Adhesion Molecules ; metabolism ; Cell Line, Tumor ; Humans ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Platelet Factor 4 ; pharmacology ; Umbilical Veins ; cytology
3.Pingyangmycin-regulated expressions of adhesion molecules in human venous malformation endothelial cells.
Yulin JIA ; Jun JIA ; Yifang ZHAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(5):760-766
Pingyangmycin (bleomycin A5 hydrochloride, PYM) is one of the anti-neoplastic agents which have been commonly used to treat venous malformations. However, the underlying mechanism by which PYM treats venous malformations remains poorly understood. It was reported that venous endothelial cells could recruit neutrophils via adhesion molecules (E-selectin, ICAM-1, ICAM-3, VCAM-1) during the acute/chronic inflammation and subsequent histological fibrosis after sclerotherapy with PYM. This study explored if the expression of E-selectin, ICAM-1, ICAM-3 and VCAM-1 in human venous malformation endothelial cells could be affected by PYM. HVMECs were cultured from human venous malformation tissue. Expressions of E-selectin, ICAM-1, ICAM-3 and VCAM-1 on HVMECs in response to PYM were analyzed by cell ELISA. The relative levels of mRNA expression in the cells were semi-quantified. The results showed that PYM up-regulated the expressions of E-selectin, ICAM-3, VCAM-1 and ICAM-1 in both time- and concentration-dependent manner. Our findings suggested that PYM could induce the expression of adhesion molecules in HVMECs, which might be a possible mechanism by which sclerotherapy by intralesional injection of PYM treats venous malformations.
Bleomycin
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analogs & derivatives
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pharmacology
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Cell Adhesion Molecules
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genetics
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metabolism
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Cells, Cultured
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Endothelial Cells
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drug effects
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metabolism
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Endothelium, Vascular
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drug effects
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metabolism
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Gene Expression
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drug effects
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genetics
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Humans
4.Effects of shear stress and lysophosphatidylcholine on adhesion molecules expression of endothelial cells.
Youqin CHEN ; Huaiqing CHEN ; Wensheng ZHANG ; Xiaoheng LIU ; Song LEI ; Yongqiu MAO
Journal of Biomedical Engineering 2002;19(4):648-672
Extensive monocyte recruitment is an early phenomenon associated with the development of atherosclerotic lesion. Although the molecular mechanisms are not completely understood, monocyte recruitment into these early lesions may involve changes in endothelial adhesion for monocyte, in which adhesion molecules expressed by endothelial cell play an active role. In vivo, the function of endothelial cells is not only affected by the chemical factors, but also by the mechanical factors. The purpose of this article was to investigate the induction of adhesion molecules expression by synergistic effects of Lysophosphatidylcholine (Lyso-PC) and shear stress in cultured human umbilical vein endothelial cells (HUVEC). The surface expression of ICAM-1, VCAM-1 and E-selectin on HUVEC induced by Lyso-PC(30 micrograms/ml) and shear stress(2.23, 4.20 dyne/cm2) were analyzed using flow cytometry. The results showed that: Compared with what were simultaneously exposed to shear stress and Lyso-PC, activating the cells with Lyso-PC prior to shear stress, or pre-conditioning the cells exposed shear stress prior to Lyso-PC incubation, a significantly higher expression of ICAM-1 and VCAM-1(P < 0.05) was resulted. HUVEC were exposed to shear stress and Lyso-PC at the same time or treated with each agonist alone, E-selectin expression was not significantly different from the control group. However, a sequential action of the two stimuli significantly increased E-selectin expression(P < 0.05). We concluded that: a sequential action of the shear stress and Lyso-PC induced an even greater expression of ICAM-1 and VCAM-1, thus it could be understood that the flows-hear stress in combination with endothelial activated by chemical factors may increase the ability of endothelial cells to recruit leukocytes even under the mechanical environment unfavorable for cell adhesion.
Cell Adhesion Molecules
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biosynthesis
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Cells, Cultured
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Endothelium, Vascular
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cytology
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drug effects
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metabolism
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Humans
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Lysophosphatidylcholines
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pharmacology
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Stress, Mechanical
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Umbilical Veins
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cytology
5.Establishment of a rat model of flutamide-induced cryptorchidism with remaining gonocytes.
Chao MA ; Da-wei HE ; Dan ZHAO ; Wen-fei HE ; Yong-bo ZHANG ; Guang-hui WEI
National Journal of Andrology 2011;17(3):208-213
OBJECTIVETo observe the effects of flutamide (Flu) on the development of testicular germocytes in SD rats, and to establish a rat model for further researches on the maldevelopment of cryptorchidism gonocytes (Go).
METHODSPregnant SD rats were subcutaneously injected with Flu from gestational day (GD) 12 to 21 to establish a model of cryptorchidism. The testes of the newborns were harvested on postnatal day (PD) 1, 10, 20 and 80 for observation of their morphological and histological changes by HE staining and detection of the expression of neural cell adhesion molecules (NCAM) by immunohistochemistry and RT-PCR.
RESULTSFlu induced 43.9% (29/66) of cryptorchidism in the exposed rats. Significant differences were found in the testicular weight and organ coefficient between the Flu and the control groups on PD 20 and 80. Gos remained in the center of seminiferous tubules of the Flu-induced testis on PD 10, and in the center of seminiferous tubules in the cryptorchids' testicular tissues on PD 20 and 80. Immunohistochemistry showed the expression of NCAM on the membrane of the remaining Gos, and RT-PCR revealed significantly up-regulated expression of NCAM mRNA in the Flu-induced testes on PD 10 and 20 as compared with the controls (P < 0.05).
CONCLUSIONA rat model of Flu-induced cryptorchidism with remaining Gos was successfully established, which could be used to study the mechanism and management of the maldevelopment of cryptorchidism gonocytes.
Animals ; Cryptorchidism ; chemically induced ; pathology ; Female ; Flutamide ; adverse effects ; Male ; Neural Cell Adhesion Molecules ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Testis ; drug effects ; metabolism ; pathology
6.Naringin inhibits monocyte adhesion to high glucose-induced human umbilical vein endothelial cells.
Ying XIONG ; Guang-fa WANG ; Jun-yan ZHANG ; Shao-yu WU ; Wei XU ; Jia-jie ZHANG ; Shu-guang WU ; Jin-jun RAO
Journal of Southern Medical University 2010;30(2):321-325
OBJECTIVETo investigate the inhibitory effect of naringin on monocyte adhesion to high glucose-induced human umbilical vein endothelial cells (HUVECs).
METHODSCultured HUVECs isolated from human umbilical cords were pretreated with or without naringin and induced with high glucose (33 mmol/L) for 48 h. Human monocyte THP-1 cells, after labeling with BCECF-AM, were co-cultured with the HUVECs for 30 min. The labeled THP-1 cells adhering to HUVECs were observed under fluoroscence microscope, and the inhibitory effect of naringin on the cell adhesion was evaluated by measuring the adhering cell density. Western blot analysis was used to detect the expressions of the adhesion molecules in the HUVECs, and reactive oxygen species (ROS) production in the HUVECs was measured using an oxidation-sensitive fluorescent probe (DCFH-DA). The nuclear extracts of the HUVECs were prepared to examine the expression of nuclear factor-kappa B (NF-kappaB) in the cell nuclei by Western blotting.
RESULTSHUVECs in high-glucose culture showed increased adhesion to THP-1 cells and enhanced expressions of the cell adhesion molecules, which were significantly attenuated by pretreatment with naringin (10-50 microg/ml). High glucose induced DCF-sensitive intracellular ROS production in the HUVECs, and this effect was inhibited by naringin pretreatment of the cells. Naringin also suppressed high glucose-induced increment of NF-kappaB expression in the cell nuclei of HUVECs.
CONCLUSIONNaringin can suppress high glucose-induced vascular inflammation possibly by inhibiting ROS production and NF-kappaB activation in HUVECs.
Cell Adhesion ; drug effects ; Cell Adhesion Molecules ; metabolism ; Cell Line ; Cells, Cultured ; Coculture Techniques ; Endothelial Cells ; cytology ; Flavanones ; pharmacology ; Glucose ; antagonists & inhibitors ; pharmacology ; Humans ; Monocytes ; cytology ; NF-kappa B ; metabolism ; Reactive Oxygen Species ; metabolism ; Umbilical Veins ; cytology
7.The effect of platelet factor 4 on the chemoattract function of the cord blood CD34+ cells.
Yi FENG ; Shi-hong LU ; Guo-li SONG ; Zhi-hua ZHANG ; Zhong-chao HAN
Chinese Journal of Applied Physiology 2003;19(2):141-144
AIMTo study the effect of PF4 and relative peptide PF4 17-70 on the chemoattract ability, the expression of adhesion molecules and CXCR4 on the flesh cord blood CD34+ cells.
METHODSCD34+ cells were separated from the cord blood using MACS immune magnetic beads, the chemoattract ability was assayed using the Transwell board, the expression of adhesion molecules and CXCR4 was measured by FACS.
RESULTS(1) PF4 and PF4 17-70 increased the migration of the CD34+ cells, the chemoattract percentage of PF4 was 157.43% +/- 50.06% (P < 0.05) and PF4 17-70 was 187.02% +/- 10.69% (P < 0.05). (2) The expression of CD49d and CXCR4 on the CD34+ cells increased after PF4 incubated, but the expressions of other adherent molecules including CD31, CD44, CD11a, CD62p, CD62E did not change.
CONCLUSIONPF4 has the chemoattract ability on the umbilical blood CD34+ cells by promoting the expression of integrin CD49d and CXCR4, PF4 may help the cord stem cells homing.
Antigens, CD34 ; metabolism ; Cell Adhesion Molecules ; metabolism ; Chemotaxis ; drug effects ; Fetal Blood ; cytology ; drug effects ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Integrin alpha4 ; metabolism ; Platelet Factor 4 ; pharmacology ; Receptors, CXCR4 ; metabolism
8.High throughput screening for intercellular adhesion molecule-1 inhibitor.
De-cheng REN ; Guan-hua DU ; Jun-tian ZHANG
Acta Pharmaceutica Sinica 2003;38(6):405-408
AIMTo develop a high throughput screening assay to identify inhibitors of intercellular adhesion molecule-1 (ICAM-1) expression in human umbilical vein endothelial cells (HUVEC).
METHODSICAM-1 expression in lipopolysaccharide-stimulated endothelial cells was measured by ELISA. The cytotoxicity of the compounds was measured by MTT.
RESULTSLipopolysaccharide (LPS) increased ICAM-1 expression in HUVEC in a concentration- and time-dependent manner. Two thousand compounds were screened and the hit rate was 1.5%. Among these 30 compounds, 24 were cytotoxic.
CONCLUSIONThe ELISA method was inexpensive, reproducible and suitable for high throughput primary cell assay. This assay was feasible to identify inhibitors of ICAM-1 and simultaneously discriminate the activity from the cytotoxic effects.
Cell Adhesion Molecules ; antagonists & inhibitors ; biosynthesis ; Cells, Cultured ; Chemistry, Pharmaceutical ; methods ; Drug Evaluation, Preclinical ; methods ; Endothelium, Vascular ; cytology ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; drug effects ; metabolism ; Lipopolysaccharides ; pharmacology ; Umbilical Veins ; cytology ; metabolism
9.Study of biological behavior of stromal cell-derived factor-1 on multiple myeloma cell migration and adhesion.
Xiao-hui ZHANG ; Jin-xiang FU ; Jian-hua ZHANG ; Yang-min ZHANG
Chinese Journal of Hematology 2006;27(4):240-243
OBJECTIVETo investigate the biological behavior of stromal cell-derived factor-1 (SDF-1) on multiple myeloma (MM) cell migration and adhesion and it related signaling pathways.
METHODSExpression of adhesion molecules on MM cells of RPMI8226, XG-1 and XG-7 cells was analysed by flow cytometry, the influence of SDF-1 on CD29 and CD49e distribution by immunofluorescence, the effect of SDF-1 on chemotaxis of MM cells by transwell assay. Activation of phosphoinositide-3 kinase (PI3K) in MM cells treated with SDF-1 and by immunoblotting.
RESULTS3 strains of MM cell line expressed many adhesion molecule. RPMI8226, XG-7 cells were all high level of expression of CD29 (> 70%). XG-1, XG-7 cells were all high level of expression of CD44 (> 80%), and XG-7 cells was of CD49d (> 90%). In all of 3 strains, the levels of expression of CD49e were low (< 30%). SDF-1 could not upregulate their expression, but could trigger the establishment of polarized morphology of MM cells and the redistribution of CD29 and CD49e. SDF-1 promoted MM cells adhesion to endothelial cells, stimulated phosphorylation of P85 subunit of PI3K in MM cells and induced MM cells migration, which were inhibited by G protein inhibitor PTX and PI3K inhibitor wortmannin.
CONCLUSIONSDF-1 can promote MM cell adhesion to endothelial cells, trigger establishment of a polarized morphology of MM cells and redistribution of adhesion molecules and induce MM cells migration via PI3K signaling pathway.
Blotting, Western ; Cell Adhesion ; drug effects ; physiology ; Cell Adhesion Molecules ; metabolism ; Cell Line, Tumor ; Cell Movement ; drug effects ; physiology ; Chemokine CXCL12 ; pharmacology ; physiology ; Enzyme Activation ; drug effects ; Flow Cytometry ; Fluorescent Antibody Technique ; Humans ; Integrin alpha4 ; metabolism ; Integrin alpha5 ; metabolism ; Integrin beta1 ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; physiopathology ; Phosphatidylinositol 3-Kinases ; metabolism ; Signal Transduction ; drug effects ; physiology
10.Morphine Attenuates Endothelial Cell Adhesion Molecules Induced by the Supernatant of LPS-Stimulated Colon Cancer Cells.
Too Jae MIN ; Sang Hee PARK ; Yi Hwa JI ; Yoon Sook LEE ; Tae Woo KIM ; Jae Hwan KIM ; Woon Young KIM ; Young Cheol PARK
Journal of Korean Medical Science 2011;26(6):747-752
A large reservoir of bacterial lipopolysaccharide (LPS) is available in the colon and this could promote colon cancer metastasis by enhancing tumor cell adhesion, intravasation, and extravasation. Furthermore, adhesion molecules like ICAM-1, VCAM-1, and E-selectin play important roles in the adhesion of tumor cells to endothelium. This study was designed to determine whether morphine can attenuate the expressions of adhesion molecules up-regulated by the supernatant of LPS-stimulated HCT 116 colon cancer cells (LPS-Sup). In this study, we divided to three groups by cell-growth medium of human umbilical vascular endothelial cells (HUVECs): the control group was incubated in growth factor-free endothelial medium, the Sup group was incubated in the supernatant of HCT 116 cells (Sup), and the LPS-Sup group was incubated in LPS-Sup. To observe effect of morphine to the adhesion molecules expressions in the LPS-Sup group, we co-treated morphine with LPS or added it to LPS-Sup. Adhesion molecule expressions on HUVECs in all three groups were measured during incubation period. Consquentially, ICAM-1, VCAM-1, and E-selectin expressions on HUVECs were significantly lower when morphine was co-treated with LPS than not co-treated. Thus, we suggest that morphine affects the expressions of adhesion molecules primarily by attenuating LPS stimuli on tumor cells.
Cell Adhesion Molecules/*metabolism
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Cell Line, Tumor
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Cell Survival/drug effects
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Colonic Neoplasms/*metabolism
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E-Selectin/metabolism
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Endothelial Cells/drug effects/metabolism
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Endothelium, Vascular/cytology
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Humans
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Intercellular Adhesion Molecule-1/metabolism
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Lipopolysaccharides/toxicity
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Morphine/*pharmacology
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Vascular Cell Adhesion Molecule-1/metabolism