Expression of integrin α5 in cervical cancer and its significance / 肿瘤研究与临床

Objective@#To investigate the expression of integrin α5 in cervical cancer, and to explore its relationship with clinicopathological characteristics of patients with cervical cancer.@*Methods@#Immunohistochemistry was used to detect the expression of integrin α 5 in cervical cancer tissues of 60 cases and normal cervical paraffin-embeded tissues of 20 cases of benign uterine lesions undergoing hysterectomy from Qingdao Municipal Hospital between January 2014 and December 2017. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expression level of integrin α5 in 20 fresh cervical cancer tissues and 20 normal cervical tissues collected from benign cervical lesions in Qingdao Municipal Hospital between January 2018 and July 2018. The relationship between the expression of integrin α5 and the clinicopathological characteristics of patients with cervical cancer was analyzed.@*Results@#The positive expression rate of integrin α5 protein in cervical cancer and normal cervical tissues was 63.3% (38/60), 35.0% (7/20), respectively, and the difference was statistically significant (χ 2 = 4.893, P < 0.05). The expression of integrin α5 mRNA in cervical cancer was 1.6±0.4 times as high as that in normal cervical tissues (t = 5.529, P < 0.01). The positive expression of integrin α5 protein was associated with lymph node metastasis in cervical cancer patients (Z = -2.636, P = 0.008).@*Conclusion@#The high expression of integrin α5 is related to lymph node metastasis of cervical cancer, and integrin α5 may be a new potential target for treatment of cervical cancer.

Expression of integrin α5 in cervical cancer and its significance / 肿瘤研究与临床

Objective To investigate the expression of integrin α5 in cervical cancer, and to explore its relationship with clinicopathological characteristics of patients with cervical cancer. Methods Immunohistochemistry was used to detect the expression of integrin α 5 in cervical cancer tissues of 60 cases and normal cervical paraffin-embeded tissues of 20 cases of benign uterine lesions undergoing hysterectomy from Qingdao Municipal Hospital between January 2014 and December 2017. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expression level of integrin α5 in 20 fresh cervical cancer tissues and 20 normal cervical tissues collected from benign cervical lesions in Qingdao Municipal Hospital between January 2018 and July 2018. The relationship between the expression of integrinα5 and the clinicopathological characteristics of patients with cervical cancer was analyzed. Results The positive expression rate of integrin α5 protein in cervical cancer and normal cervical tissues was 63.3％(38/60), 35.0％(7/20), respectively, and the difference was statistically significant (χ2=4.893, P<0.05). The expression of integrin α5 mRNA in cervical cancer was 1.6 ±0.4 times as high as that in normal cervical tissues (t= 5.529, P< 0.01). The positive expression of integrin α5 protein was associated with lymph node metastasis in cervical cancer patients (Z= -2.636, P= 0.008). Conclusion The high expression of integrinα5 is related to lymph node metastasis of cervical cancer, and integrin α5 may be a new potential target for treatment of cervical cancer.

Expression and significance of integrin α5β1 and fibronectin in atherosclerotic plaques from autopsy specimens / 中华病理学杂志

Objective@#To investigate the expression of integrin α5β1 and fibronectin in the human aorta and coronary artery, and their effects in the development of atherosclerosis.@*Methods@#One hundred and twenty autopsy aorta and coronary artery specimens were collected, and the expression of CD68, actin, integrin α5β1 and fibronectin was detected by immunohistochemical staining. Atherosclerotic plaques were located by CD68 and actin staining, and the degree of coronary artery stenosis was determined by elastic fiber staining and NIH Scion Image(60.1) software. The coronary artery tissues were divided into groups A (0-25%); B (26%-50%); C (51%-75%) and D (76%-100%) according to the degree of stenosis.@*Results@#Integrin α5β1 showed cytoplasmic expression in endothelium, foam cells, monocytes, smooth muscle cells and adjacent tissue around calcification. In both the aorta and coronary artery, integrin α5β1 expression was stronger in the smooth muscle cells in the internal elastic lamina than in the tunica. The expression intensity in coronary artery smooth muscle decreased with increasing degree of coronary artery stenosis. Fibronectin showed cytoplasmic expression in foam cells, monocytes, smooth muscle cells of the internal elastic lamina and adjacent tissue around calcification. There was positive correlation of fibronectin and integrin α5β1 expression in smooth muscle cells and adjacent tissue around calcification.@*Conclusions@#In the development of atherosclerosis, integrin α5β1 and fibronectin may participate in the regulating the migration of smooth muscle cells to the intima, and promote the formation of local calcification of atherosclerotic plaques. But integrin α5β1 is not involved in the late stage of atherosclerosis with increasing coronary artery stenosis.

Heat or radiofrequency plasma glow discharge treatment of a titanium alloy stimulates osteoblast gene expression in the MC3T3 osteoprogenitor cell line

PURPOSE: The purpose of this study was to determine whether increasing the Ti6Al4V surface oxide negative charge through heat (600degrees C) or radiofrequency plasma glow discharge (RFGD) pretreatment, with or without a subsequent coating with fibronectin, stimulated osteoblast gene marker expression in the MC3T3 osteoprogenitor cell line. METHODS: Quantitative real-time polymerase chain reaction was used to measure changes over time in the mRNA levels for osteoblast gene markers, including alkaline phosphatase, bone sialoprotein, collagen type I (alpha1), osteocalcin, osteopontin and parathyroid hormone-related peptide (PTH-rP), and the osteoblast precursor genes Runx2 and osterix. RESULTS: Osteoprogenitors began to differentiate earlier on disks that were pretreated with heat or RFGD. The pretreatments increased gene marker expression in the absence of a fibronectin coating. However, pretreatments increased osteoblast gene expression for fibronectin-coated disks more than uncoated disks, suggesting a surface oxide-mediated specific enhancement of fibronectin's bioactivity. Heat pretreatment had greater effects on the mRNA expression of genes for PTH-rP, alkaline phosphatase and osteocalcin while RFGD pretreatment had greater effects on osteopontin and bone sialoprotein gene expression. CONCLUSIONS: The results suggest that heat and RFGD pretreatments of the Ti6Al4V surface oxide stimulated osteoblast differentiation through an enhancement of (a) coated fibronectin's bioactivity and (b) the bioactivities of other serum or matrix proteins. The quantitative differences in the effects of the two pretreatments on osteoblast gene marker expression may have arisen from the unique physico-chemical characteristics of each resultant oxide surface. Therefore, engineering the Ti6Al4V surface oxide to become more negatively charged can be used to accelerate osteoblast differentiation through fibronectin-dependent and independent mechanisms.

T-CAM, a fastatin-FIII 9-10 fusion protein, potently enhances anti-angiogenic and anti-tumor activity via alphavbeta3 and alpha5beta1 integrins

We made fusion protein of fastatin and FIII 9-10, termed tetra-cell adhesion molecule (T-CAM) that can interact simultaneously with alphavbeta3 and alpha5beta1 integrins, both playing important roles in tumor angiogenesis. T-CAM can serve as a cell adhesion substrate mediating adhesion and migration of endothelial cells in alphavbeta3 and alpha5beta1 integrin-dependent manner. T-CAM showed pronounced anti-angiogenic activities such as inhibition of endothelial cell tube formation, endothelial cell proliferation, and induction of endothelial cell apoptosis. T-CAM also inhibited angiogenesis and tumor growth in mouse xenograft model. The anti-angiogenic and anti-tumoral activity of molecule like fastatin could be improved by fusing it with integrin-recognizing cell adhesion domain from other distinct proteins. The strategy of combining two distinct anti-angiogenic molecules or cell adhesion domains could facilitate designing improved anticancer agent of therapeutic value.

Integrin alphavbeta3, alpha5beta1 Effects on Cell Proliferation and Migration in Human Osteosarcoma / 대한정형외과연구학회지

BACKGROUND: We investigate the influence of cell surface adhesion receptor integrin alphavbeta3, alpha5beta1 contributes to proliferation and migration of tumor cell in osteosarcoma for carves out a new treatment model by regulation of integrin roles in human osteosarcoma. MATERIALS AND METHODS: We performed proliferation assay, total 11 cell lines including 7 osteosarcoma cell lines established from patients and 4 osteosarcoma standard cell lines. Murine monoclonal anti-alpha5beta1 and anti-alphavbeta3 (Chemicon International Inc. Temecula, CA) were used for growth inhibition assays. We also performed cell motility assay by using the Boyden chamber to evaluate the effect of integrin mediated cell migration. We used the HOS standard osteosarcoma cell lines and each separates contained serum free media with mouse IgG1 negative control antibody, anti-alpha5beta1 antibody and anti-alphavbeta3 antibody. RESULTS: Proliferation of cells decreased significantly in 10 out of 11 cell lines when blocking with alphavbeta3 or alpha5beta1 respectively. Blocking with anti-alphavbeta3 antibody decreased significantly tumor cell proliferation in 10 cell lines. Among the 10 cell lines, 7 cell lines showed significantly more decrease of proliferation with anti-alphavbeta3 antibody than with anti-alpha5beta1antibody. Blocking with anti-alpha5beta1 antibody decreased significantly tumor cell proliferation in 10 cell lines. Among the 10 cell lines, 3 cell lines showed significantly more decrease of proliferation with anti-alpha5beta1 antibody than with anti-alphavbeta3 antibody. Including statistically not significant 2 cell lines the growth inhibition of osteosarcoma cell lines was more obvious (10 out of 11) in blocking with anti-alphavbeta3 antibody. The migration of cells was significantly decreased when blocked with anti-alpha5beta1 antibody and anti-alphavbeta3 antibody. CONCLUSION: Under the based on the integrin alphavbeta3, alpha5 beta1 are central role on proliferation and migration of osteosarcoma cells, we could be more approach to new therapeutic endeavors with antibody to integrin alphavbeta3, alpha5beta1 molecular target of osteosarcoma.

Effect of Recombinant Fragment of Fibronectin on the Cellular Functions of Human Bone Marrow-Derived Mesenchymal Stem Cells / 대한정형외과연구학회지

PURPOSE: We investigated the effects of recombinant 9-10th type III repeat of fibronectin (rhFNIII9-10) on the adhesion, proliferation, and the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells(hMSCs). MATERIALS AND METHODS: Adhesion and blocking assay for hMSCs were performed on the plates which had been coated with 100 microgram/ml rhFNIII9-10 or fibronectin. hMSCs seeded on the precoated plates were cultured in the osteogenic media for 3 weeks. MTS(Dimethylthiazole carboxymethoxyphenyl sulfophenyl tetrazolium compound) assay for the cell number, [Methyl-3H] thymidine incorporation study, alkaline phosphatase activity assay, calcium content assay and RT-PCR for alkaline phosphatase, osteopontin, cbfa-1, and type I collagen were performed during the osteogenic differentiation. RESULTS: hMSCs showed significantly increased adhesion to rhFNIII9-10-coated plates and fibronectin-coated plates. A monoclonal antibody to the integrin alpha 5 beta 1 inhibited adhesion to rhFNIII9-10-coated plates and fibronectin-coated plates in dose-dependent manner. hMSCs seeded on the rhFNIII9-10-coated plates showed increased proliferation during the osteogenic differentiation. However, there was no significant difference in the alkaline phosphatase activity, calcium content and expression levels of mRNAs for alkaline phosphatase, osteopontin, cbfa-1, and type I collagen of hMSCs seeded on the rhFNIII9-10-coated plates. CONCLUSION: rhFNIII9-10 stimulates hMSCs adhesion and increases hMSCs proliferation during the osteogenic differentiation. Although osteogenic differentiation is not promoted, adsorption of rhFNIII9-10 onto appropriate biomaterials can enhance integrin-mediated hMSCs adhesion and proliferation. This biomolecular engineering strategy represents a robust approach to increase biofunctional activity and integrin specificity of hMSCs.

Relation of ME491/CD_(63) gene and integrin ?5 in the invasion and metastases of ovarian cancer / 中华妇产科杂志

0.05). There was significant difference between G_1, G_2and G_3(P0.05). Low expression levels in G_4 (Ⅲ,Ⅳ) were observed.Significant differences were noted between expression levels in G_4 (Ⅲ,Ⅳ) and in G_2, G_3 or G_4 (Ⅰ,Ⅱ; P