ABSTRACT
Focal adhesion kinase (FAK) plays a critical role in the process of cell adhesion and migration by regulating the expression of downstream small G proteins. A kind of focal adhesion kinase (FAK) inhibitor was used to inhibit the phosphorylation of Y397 site of FAK, and scratch wound migration assay was used to examine the effect of FAK inhibitor with different concentrations (0-250 nmol/mL) on the migration of hepatomal cells (Hep G2 cells) at 0, 2, 4, 8 and 24h. Immunofluorescence analysis and Western blot analysis were performed to detect the expression of F-actin and small G proteins Rac1, RhoA and Cdc42 in Hep G2 cells treated with FAK inhibitor for 120 min. The results indicated that the FAK inhibitor can inhibit the migration of Hep G2 cells with a dose- and time-dependent manner. F-actin was down-regulated in Hep G2 cells treated with FAK inhibitor for 120 min, and expression of small G proteins were inhibited at different durations. The inhibition of FAK phosphorylation could inhibit cell adhesion and migration by down-regulating small G proteins. These results suggested that FAK inhibitor can inhibit the migration of tumor cells by blocking FAK phosphorylation. This means that FAK inhibitor can block the metastasis of tumor cells to surrounding tissues. It may be a potential application in the prevention and treatment of cancer.
Subject(s)
Humans , Cell Adhesion , Cell Movement , Focal Adhesion Protein-Tyrosine Kinases , Metabolism , Physiology , Hep G2 Cells , Liver Neoplasms , Pathology , Neoplasm MetastasisABSTRACT
Tumor angiogenesis induced by vascular endothelial cells (VECs) migration is a necessary condition for tumor growth and metastasis. The purpose of this study is to investigate the effect of focal adhesion kinase (FAK) inhibitor (50nmol/mL) on the adhesion and migration of endothelial cells(ECs) and the expression of focal adhesion proteins vinculin, talin and paxillin. Scratch wound migration assay was performed to examine the effect of FAK inhibitor with 50nmol/mL on ECs migration at 0, 5, 10, 30, 60 and 120min, respectively. And immunofluorescence analysis was performed to detect the expression of F-actin in ECs treated with FAK inhibitor within 2h. Western blot was carried out to determine the effect of FAK inhibitor on expression of vinculin, talin and paxillin proteins. The results showed that the migration distance and the expression of F-actin in ECs treated with FAK inhibitor decreased significantly compared with that of the controls, and the level of vinculin showed no significant difference with increasing of treated time of FAK inhibitor. However, the talin and paxillin showed an identical decreasing tendency in 5-10min, but slowly going up in 30min and then after subsequently decreasing. The results of this study proved that blocking phosphorylation of FAK could inhibit VECs adhesion and migration by downregulating focal adhesion proteins so that it may inhibit tumor angiogenesis. This may provide a new approach for tumor therapy.
Subject(s)
Humans , Cell Adhesion , Cell Movement , Physiology , Cells, Cultured , Endothelial Cells , Cell Biology , Metabolism , Focal Adhesion Protein-Tyrosine Kinases , Metabolism , Focal Adhesions , Metabolism , Physiology , Neoplasms , Neovascularization, Pathologic , Paxillin , Metabolism , Talin , Metabolism , Vinculin , MetabolismABSTRACT
Aseptic loosening is mainly mediated by bone resorption cytokines in the surrounding area of orthopaedic implants. Our previous investigation demonstrtated that different-sized titanium particles loading can inbibit the osteoblastic differentiation and mineraliztion. In order to investigate the hypothesis that particulate wear debris derived from prosthetic biomaterials affects the release of bone resorption related cytokines, we studied the influence of different-sized titanium particles loading on the osteoblastic cytokines by assaying the secretion of IL-6, IL-10 with use of ABC-quantitative sandwich enzyme-linked immunosorbent assay, and on the expression of osteoclast differentiation factors (ODF) by RT-PCR. The results showed that the 0.9 microm titanium particles promoted osteoblasts producing bone resorption cytokines (IL-6, ODF), and simultaneously secreted bone absorption restraining factor (IL-10) quickly and transitorily. In comparison, the 2.7 microm and 6.9 microm titanium particles,especially the latter primarily promoted osteoblasts secreting bone absorption promoting factors powerfully and slowly. The results suggested that there was a biphasic response appearing in titanium particles loaded-osteoblastic cultures, the level of which varied according to the different size and the loading time of titanium particles. This in vitro experimental result showed that attentaion to the inhibition of bone resorption cytokines stimulated by wear debris and to the screen potential favourable biomaterials for implants must be taken.
Subject(s)
Animals , Female , Male , Rabbits , Cells, Cultured , Cytokines , Bodily Secretions , Interleukin-10 , Bodily Secretions , Interleukin-6 , Bodily Secretions , Joint Prosthesis , Osteoblasts , Metabolism , Particle Size , Prosthesis Failure , RANK Ligand , Bodily Secretions , Titanium , PharmacologyABSTRACT
300 cases of atrophic gastritis were typed into 3 groups according to TCM principle, viz. Qideficiency of the Spleen-Stomach, Yang-deficiency of the Spleen-Stomach and Yin-deficiency of the Spleen-Stomach. Biopsy specimens were examined pathologically and detection of Helicobactor pylori performed for comparism. Results showed that in the Yin-deficiency type, the severity of lesion in the gastric mucosa and its type of lesion, severity of atrophy of mucosal glands, infiltration of interstitial inflammatory cells were most serious, while for the detection of Helicobactor pylori, it made no difference among all the three types.