Overexpressions of Vimentin and Integrins in Human Metastatic Spine Tumors

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.

Study on the specific immunity induced by dendritic cell vaccine loading allogenic microvascular endothelial cell bEnd. 3 antigen against U14 cervical cancer cell in mice / 中华妇产科杂志

Objective To explore the specific cellular and humoral immunity induced by dendritic cells (DC) vaccine loading allogenic microvascular endothelial cell bEnd. 3 antigen against U14 cervical cancer cell of mice. Methods Mouse brain microvascular endothelial cell bEnd. 3 was cultured and identified for preparation endothelial cell bEnd. 3 antigen. The level of mRNA expression of vascular endothelial growth factor receptor 2 (VEGF-R2) and integrin αV was detected by reverse transcription (RT)-PCR. The BALB/c mice were immuned with DC loading bEnd. 3 antigen 4 times in 4 weeks (bEnd. 3-DC group), while the mice only were immuned with DC or injected with phosphate buffer saline (PBS group) as control group. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, cytotoxic T lymphocyte (CTL) response of spleen lymphocytes in vitro, the percentage of CD3+ CD+8 surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and western blot. Results The expression of VEGF-R2 and integrin αV gene in bEnd. 3 cells were expressed highly.After the vaccine was injected, the tumors of mice in PBS group grew faster than those in other groups, while the tumors in bEnd. 3-DC group grew slowly and disappeared after 2 weeks. The volume of tumors in DC group grew slower than those in PBS group [(0.11± 0.13) cm3 versus (3.38 ±0.34) cm3]. The CTL response of spleen iymphocytes in vitro showed that bEnd. 3-DC cells could kill bEnd. 3 cells, the special lysis rate was more than 60% . The percentage of CD+3 CD+8 spleen lymphocytes in bEnd. 3-DC group[(38.6 ± 0.7) %] was higher than those in other groups (P ＜ 0.05). The titer of serum antibody of Immunocytochemistry analysis indicated there were specific antigen-antibody reaction to bEnd. 3 cell in bEnd. 3-DC group. Western blot analysis revealed that there were specific bands at 220 000 (VEGF-R2).Conclusions bEnd. 3-DC vaccine can inhibit the tumor growth of U14 cervical cancer cell of mice, which indicates that the special cellular and humorai immunity are induced by bEnd. 3-DC antigen which maybe have some antigens in bEnd. 3 cells that reacts with endothelial cell proliferation-related antigens.

Inhibitory Effect of Antibody to alphavbeta5 in Corneal Angiogenesis

PURPOSE: This study investigated the importance of alphavbeta5 function during vascular endothelial growth factor (VEGF) induced corneal angiogenesis by examining the effects of antibody to alphavbeta5 that blocks alphav 5-mediated cell adhesion to vitronectin. METHODS: A hydrogel disk containing 500 ng of VEGF was implanted into the superior corneal stroma of each of sixteen New Zealand white rabbit eyes. Each eye also received a second hydrogel disk placed adjacent to the first, randomized to contain either 40 g of antibody to alphavbeta5 (n=8) or phosphate-buffered saline (PBS)(n=8). Both disks were positioned 1.2 mm apart from the superior limbus. Eyes were examined daily under a stereomicroscope by two observers and assigned an angiogenesis score based on number and length of new blood vessels. RESULTS: On days 3 through 7 postimplantation, angiogenesis scores were significantly lower in eyes treated with antibody to alphavbeta5 (averaged score=16.33) as compared to eyes treated with PBS (averaged score=26.52)(P<0.05, Wilcoxon signed rank test). CONCLUSIONS: In a rabbit corneal micropocket assay, antibody to alphavbeta5 inhibits corneal angiogenesis induced by VEGF. Substances that target the integrin alphavbeta5 subunit may have therapeutic potential in disorders characterized by ocular neovascularization.