OBJECTIVETo investigate the expression and distribution pattern of Integrin alphaV beta3 (ItgalphaV beta3) in course of distraction 44 adults New Zealand white osteogenesis and to quest for the function of ItgalphaV beta3 in distraction osteogenesis (DO).

METHODSrabbits were selected and divided randomly into 3 groups: DO, bone fracture and normal group. There were 20, 20 and 4 rabbits in each group separately. Animal models were established and animals were killed at different time points. Sections were stained by IHC method. Distribution and expression pattern of ItgalphaV beta3 were observed by semi-quantitative technique, and the results were managed with statistic methods.

RESULTSThe expression of ItgalphaV beta3 was found both in the DO and bone fracture groups. The positive staining was seen mainly in vascular endothelial cells on cytomembrane and in cytoplasm. The staining intensity of group of DO was higher than that of the bone fracture group.

CONCLUSIONItgalphaV beta3 plays an important role in promoting the process of DO.

OBJECTIVETo explore the effect of zoledronate (ZOL) on the osteoclast adhesion and expression of integrin α(v) and β3 in vitro.

METHODSMice RAW264.7 cells were used for osteoclast differentiation in vitro, and osteoclastogenesis was examined by tartrate-resistant acid phosphatase (TRAP) staining and dentin resorption lacunae examination. The cells were then divided into 2 groups, the control group and ZOL treatment group (treated with 1 x 10(-6) mol · L(-1) ZOL for 2 d). The adhesion ability of osteoclasts and mRNA and the protein expressions of integrin α(v) and β3 were examined by crystal violet staining, real-time fluorescence quantitative polymerase chain reaction, Western blot analysis, and immunofluorescent chemistry.

RESULTSTRAP staining and dentin resorption lacunae examination revealed the formation of multi-nuclear osteoclasts. ZOL treatment significantly decreased the adhesion ability of osteoclasts (P < 0.01). In the ZOL-treated group, the mRNA levels of integrin α(v) and β3 were 0.66 ± 0.05 and 0.59 ± 0.08, respectively. In the control group, the mRNA levels of integrin α(v) and β3, were 1.01 ± 0.01 and 1.01 ± 0.02, respectively; these values were higher than those in the ZOL-treated group (P < 0.01). The protein level of integrin α(v) and β3 in the ZOL-treated group (31,934.84 ± 112.91 and 18,812.79 ± 194.13) was downregulated by approximately 39.19% and 40.17%, respectively, compared with those in the control group (52,517.81 ± 211.72 and 31,441.93 ± 456.87) (P < 0.01). Immunofluorescent examination showed that the fluorescent intensities of integrin α(v) and β3 in the ZOL-treated group (9.491 ± 0.748 and 4.744 ± 0.759) were also significantly decreased compared with those in the control group (15.159 ± 1.143 and 11.418 ± 1.095) (P < 0.01).

CONCLUSIONZOL significantly inhibits osteoclast adhesion and downregulates integrin α(v) and β3, expression, thus contributing to the ZOL-induced inhibition of osteoclast- mediated bone resorption.

BACKGROUND: It has been known that integrins are not only simple glue which mediate cell-to-cell or cell to extra-cellular matrix but also function as signaling molecules at the surface of cell by conformational change. Because of the diversity of subunits and versatility of ligand recognition, integrins have been recognized as important molecules in kidney and the expression of integrin in normal human kidney has already been reported. METHODS: To know the significance of integrin expression in the pathogenesis of various renal diseases, we stained kidney specimens from the patients with membrano- proliferative glomerulonephritis(MPGN), minimal change nephrotic syndrome(MCNS) and membraneous nephropathy(MN) of lupus nephritis with 13 monoclonal antibodies of integrins. RESULTS: There was a significant change in the distribution of integrin subunits according to the types of glomerular diseases. Integrin subunit alpha2 was helpful in confirming the mesangial interposition as there was intense staining of subendotherial area in MPGN, while only the mesangium was stained with this unit in normal. In addition, an abundant staining of alpha4 and beta2 was observed and this finding indicated non-glomerular resident cell is participating in the MPGN, while in MN type of lupus nephritis, alpha2 staining was restricted to mesangium and alpha4 or beta2 integrin staining was negative. In the MN type of lupus nephritis, the remarkable finding was a ragged appearance in the subepithelial area, just below the glomerular basement membrane, which was visible in alpha3 and alphaV integrin staining. In MCNS, there was no difference in the staining pattern of integrin. CONCLUSION: Integrin subunits can be useful in evaluating the specific cell types which is actively involved in pathogenesis and these results suggest that alteration of integrin distribution can play an important role in the pathogenesis of glomerular diseases.
Adhesives ; Antibodies, Monoclonal ; Antigens, CD18 ; Glomerular Basement Membrane ; Glomerulonephritis ; Glomerulonephritis, Membranoproliferative ; Humans ; Integrin alphaV ; Integrins ; Kidney ; Lupus Nephritis

Liver fibrosis is a pathological process of the excessive accumulation of extracellular matrix, especially collagen al (I) in liver. Ultimately, hepatic fibrosis leads to cirrhosis or hepatic failure. Liver fibrosis and early cirrhosis can be reversed, thus control of the development of liver fibrosis is very important for preventive treatment of cirrhosis and hepatic failure. This is a review of potential targets for anti-hepatic fibrosis based on plenty of publications, including TGF-β1 and integrin α(v) and so on, aimed at providing novel therapeutic targets in liver fibrosis.
Collagen ; metabolism ; Humans ; Integrin alphaV ; metabolism ; Liver ; pathology ; Liver Cirrhosis ; drug therapy ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVE: To comparatively investigate the expression of several integrins in specimens of human bone metastases and degenerative bone tissue. METHODS: Degenerative cancellous tissue was obtained from a sample of human degenerative spine. Thirteen human specimens were obtained from metastatic spine tumors, whose primary cancer was colon cancer (n=3), hepatocellular cancer (n=3), lung cancer (n=4), and breast cancer (n=3). The expression of vimentin and integrins alphav, beta1, and beta3 was assessed in metastatic and degenerative specimens by immunohistochemistry and real-time reverse transcription-polymerase chain reaction (qRT-PCR). RESULTS: Immunohistochemical staining showed that vimentin and integrin alphav was broadly expressed in all tissues examined. By contrast, integrin beta1 was weakly expressed only in 38.4% (5/13) of tissues. Integrin beta3 was consistently negative in all cases examined. qRT-PCR analysis showed that vimentin gene expression was higher in all metastatic specimens, as compared to degenerative bone. The gene expression of integrin alphav in breast specimen was significantly higher than others (p=0.045). The gene expression of integrin beta1 was also higher in all metastatic specimens than in degenerative bone tissue. The gene expression of integrin beta3 was variable. CONCLUSION: Spinal metastatic tumors have mesenchymal characteristics such as increased expression of vimentin. The increased expression of integrin alphav and beta1 in spine metastatic tumors suggests that adhesive molecules such as integrin may have implications for the prevention of spine metastasis.
Adhesives ; Antigens, CD29 ; Bone and Bones ; Breast ; Breast Neoplasms ; Colonic Neoplasms ; Gene Expression ; Humans ; Immunohistochemistry ; Integrin alphaV ; Integrin beta3 ; Integrins* ; Liver Neoplasms ; Lung Neoplasms ; Neoplasm Metastasis ; Spine* ; Vimentin*

BACKGROUND AND OBJECTIVES: Integrins mediate the migration, adhesion and proliferation of vascular smooth muscle cells. Adenosine diphosphate (ADP) can activate vascular integrins. We assessed the hypothesis that 'statins inhibit the ADP-stimulated activation of integrins alpha v beta5 and alpha v beta3 in human aortic smooth muscle cells (HASMC)'. MATERIALS AND METHODS: The expressions of integrins were analyzed using flow cytometry. The activations of integrins were evaluated using the adhesion assay, with prothrombin as an activation-dependent ligand. The MTT assay was used to evaluate the proliferation. RESULTS: Statins did not suppress the expressions of the integrins, alpha v beta5 and alpha v beta3. The ADP-stimulated adhesion was partially prevented by LM609, which blocked integrin alpha v beta3 (13% inhibition), and markedly prevented by P1F5, which blocked integrin alpha v beta5 (76% inhibition; n=5, p<0.05). However, the proliferation was inhibited by c7E3 and LM609, but not by P1F5. Statins inhibited the ADP-stimulated adhesions in a dose-dependent manner after 15 min of pretreatment. After incubating HASMC with statins for 1 day, simvastatin and fluvastatin inhibited the adhesion by 70 and 66%, respectively (n=5, p<0.05 vs. no statin). Statins also inhibited the ADP-stimulated proliferation of HASMC. CONCLUSION: Statins did not suppress the expressions of the integrins, alpha v beta5 and alpha v beta3, but did inhibit the ADP-stimulated activation of the integrins of HASMC.
Adenosine Diphosphate ; Flow Cytometry ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors* ; Integrin alpha Chains ; Integrin alphaV ; Integrins* ; Muscle, Smooth, Vascular* ; Myocytes, Smooth Muscle ; Prothrombin ; Simvastatin

Netrins are secreted molecules and involved in axon guidance, cell migration and tumor development. Recent studies revealed that netrins perform novel functions in such processes as epithelial development and angiogenesis without operating through the classical netrin receptors, DCC (Deleted in Colorectal Cancer) and Unc5h. In the present study, we investigated the roles of netrin-1 and its receptors in cell spreading of human glioblastoma cells, and found that netrin-1 haptotactically enhanced fibronectin-induced cell spreading and focal adhesion formation in U373 glioblastoma cells. Netrin-1 binding to the U373 cell membrane was blocked by an antibody against alpha v integrin subunit, but not by an anti-DCC or anti-Unc5h antibody. In addition, enhancement of the fibronectin response by netrin-1 was abrogated by a function blocking antibody against integrin alpha v beta 3. Since the alpha v subunit of the integrin family plays an important role in the pathophysiological aspects of cell migration, including tumor angiogenesis and metastasis, our data provide important insight into the molecular mechanism of netrin function.
Axons ; Cell Membrane ; Cell Movement ; Fibronectins ; Focal Adhesions ; Glioblastoma ; Humans ; Integrin alphaV ; Integrin alphaVbeta3 ; Neoplasm Metastasis ; Nerve Growth Factors ; Receptors, Cell Surface ; Tumor Suppressor Proteins

INTRODUCTION: The pathophysiology of PIH remains unclear. Recently, placental abnormalitiesare stressed as a possible cause of PIH. Abnormal shallow invasion of trophoblasts, confinedto decidua, without involving myometrium is believed to result in reduced uteroplacentalperfusion, endothelial injury, and activation of coagulation cascade system. Integrin, one of theadhesive membrane proteins, is expected to be related to the regulation of trophoblasts invasion. PURPOSE: The purpose of this study is to investigate the expression of adhesion moleculesin placenta and the correlation between uterine artery Doppler findings and integrinexpressions in the placentas of PIH patients. SUBJECTS: Thirty-six cases of severe PIH patients were enrolled in the study with 10number of normal control pregnant women. The integrin subunit expressions withimmunohistochemical staining were observed in floating villi, maternal-side cytotropholbasts, andfetal-side cytotrophoblasts. Uterine artery Doppler study was also performed, and the S/Dratio was evaluated. Abnormal Doppler findings was defined as S/D ratio>or=2.6. RESULTS: Cytoplasmic staining of villi and placental bed cytotrophoblast for theintegrin alpha1 subunit in PIH specimen was weaker than those in normal controls. Theexpression of integrin beta1 subunit was negative for both controls and PIH group. Thepositive cytoplasmic stain was observed among PIH placenta in contrast to normal control inwhich the expression of integrin beta4 subunit was not detected. The expression of alpha v beta3 introphoblast with PIH was positive staining, but not in control group. Uterine artery Dopplervelocimetry was performed in 25 cases with PIH. Trace(+/-) or - staining of integrin alpha1 subunit were observed in 60.0% of abnormal S/D(>or=2.6) group, 20.0% of normal S/Dratio group patients, respectively. Trace or + staining of integrin beta4 subunit were observedin 50.0% of abnormal S/D group and 6.7% of normal S/D group and this is in statisticallysignificant. Trace or + staining of integrin alpha v beta3 subunit were observed 70.0% ofabnormal S/D group and 26.7% of normal S/D group, and this statistically significant. CONCLUSION: In PIH the abnormality in the invasion of cytotrophoblats results inabnormal integrin subunit expression, but it is also correlated to the abnormal uterine arteryDoppler velocimetry which shows a S/D ratio of greater than 2.6. Thus, the uterine arteryDoppler velocimetry reflects abnormal placentation.
Animals ; Antigens, CD29 ; Cytoplasm ; Decidua ; Female ; Humans ; Hypertension, Pregnancy-Induced* ; Integrin alpha1 ; Integrin alphaV ; Integrin beta4 ; Integrins ; Membrane Proteins ; Mice ; Myometrium ; Placenta* ; Placentation ; Pregnancy ; Pregnant Women ; Rheology* ; Trophoblasts ; Uterine Artery*

OBJECTIVE: This study was to investigate the expression and significance of Integrins subunits in laryngeal squamous cell carcinoma (LSCC). METHOD: The expression of Integrins subunits was detected by cDNA microarray in 4 cases of primary LSCC tissues and corresponding adjacent normal tissues. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) were used to identify the different expression of Integrins subunits in 24 cases of primary LSCC tissues and corresponding adjacent normal tissues. RESULT: A cDNA microarray analysis revealed significant changes in the expression of Integrins subunits, with IntegrinalphaV, Integrinbeta8 being up-regulated and Integrinalpha8 being down-regulated. The result of RT-PCR was consistent with that of cDNA microarray. The mRNA levels of IntegrinalphaV and Integrinbeta8 were significantly higher in LSCC tissues than that in corresponding adjacent normal tissues (1.0131 +/- 0.4780 vs 0.7591 +/- 0.4678 for IntegrinalphaV, P<0.05, 1.7362 +/- 1.3849 vs 1.2267 +/- 0.9363 for Integrinbeta8, P<0.05). The mRNA levels of Integrinalpha8 were significantly lower in LSCC tissues than that in corresponding adjacent normal tissues (0.2646 +/- 0.2622 vs 0.5457 +/- 0.3827, P<0.05). CONCLUSION The expression of IntegrinalphaV, Integrinbeta8, Integrinalpha8 were significantly up-regulated or down-regulated in laryngeal squamous cell carcinoma, which may relate to tumorigenesis and development of laryngeal squamous cell carcinoma.
Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Humans ; Integrin alpha Chains ; genetics ; metabolism ; Integrin alphaV ; genetics ; metabolism ; Integrin beta Chains ; genetics ; metabolism ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Neoplasm Staging

In postmenopausal osteoporosis, estrogen deficiency leads to unbalance of bone metabolism, decreased bone formation and increased bone resorption, and the result is reduced bone mineral density (BMD) and bone stiffness. The processes of bone formation and resorption involves the expression of integrins in anchoragedependent cells, such as osteoblast and osteoclast. The osteoporosis-induced rats frequently demonstrated the loss of trabecular bone volume in the tibia, vertebra and mandible due to estrogen depletion. However, in maxilla, study has been rare because of its anatomical limits. So the objective of this study was to investigate bony change and property of integrin expression in maxilla of osteoporosis-induced rats. 12-week-old female Sprague-Dawley rats were bilaterally ovariectomized (OVX). At 1, 2, 3, 4, 8 and 12 weeks, control and OVX group rats were sacrificed respectively. BMD of maxilla of the rats was measured using dual-energy X-ray absorptiometry (DEXA). And then the histopathologic observation, histomorphometric analysis and immunohistochemistry with CD44, alpha2 integrin, alpha5 integrin, alpha6 integrin, alphav integrin and beta3 integrin were done. BMD of alveolar bone in maxilla was decreased with significance statistically after OVX 4 weeks and was decreased 18.15% at OVX 12 weeks group compared to control group. From OVX 4 to 12 weeks, the thickness of periodontal ligament space was decreased, the number of osteoclast and the size of marrow stroma were increased than control group. By histomorphometric analysis, the size of marrow stroma of alveolar bone in maxilla was increased 86.42% at OVX 12 weeks group compared to control group. CD44 was widely expressed throughout the odontoblast, cementoblast, dental pulp, preiodontal ligament, osteocyte, osteoclast and perivascular tissue at control group, and CD44 immunoreactivity was increased the odontoblast, osteoblast and osteoclast at OVX groups. alpha2 integrin was expressed the odontoblast and osteoblast at control group, but alpha2 integrin immunoreactivity was decreased the osteoblast at OVX 12 weeks group. alpha5 integrin was expressed the cementoblast, osteoblast and osteoclast at control group, and alpha5 integrin immunoreactivity was decreased the osteoblast and was increased the osteoclast from OVX 4 weeks group. alpha6 integrin was weakly expressed the odontoblast, cementoblast, osteoblast and osteoclast at control group, and alpha6 integrin immunoreactivity was weakly increased the osteoclast from OVX 4 weeks. alphav integrin was expressed the odontoblast and osteoclast at control group, and alphav integrin immunoreactivity was strongly increased the osteoclast from OVX 4 weeks. beta3 integrin was expressed the osteocyte and osteoclast at control group, and beta3 integrin immunoreactivity was strongly increased the osteoclast from OVX 4 weeks. From these results, alveolar bone in maxilla of OVX rats was decreased BMD gradually. Moreover, alpha2 and alpha5 integrin expression of osteoblast was decreased, and alpha5, alphav and beta3 integrin expression of osteoclast was increased in OVX rats. Thus, this study indicates that consideration of reduced BMD is necessary in dental procedure of postmenopausal women.
Absorptiometry, Photon ; Animals ; Bone Density* ; Bone Marrow ; Bone Resorption ; Dental Cementum ; Dental Pulp ; Estrogens ; Female ; Humans ; Immunohistochemistry ; Integrin alpha2 ; Integrin alpha5 ; Integrin alpha6 ; Integrin alphaV ; Integrin beta3 ; Integrins ; Ligaments ; Mandible ; Maxilla* ; Metabolism ; Odontoblasts ; Osteoblasts ; Osteoclasts ; Osteocytes ; Osteogenesis ; Osteoporosis, Postmenopausal ; Ovariectomy* ; Periodontal Ligament ; Rats* ; Rats, Sprague-Dawley ; Spine ; Tibia