Methods: (1) Nude mice bearing tumor xenografts of human colon carcinoma were injected intravenously with 18.5 MBq 99Tcm-GX1, and ECT imaging were performed; (2) Immunohistochemistry and immunofluorescence were performed to evaluate the binding ability of GX1 to RMEC; (3) Antiangiogenesis ability of GX1 on RMEC were analyzed by in vitro MTT assay, migration assay, and tube formation assay. Results: (1) ECT imaging indicated that tumor on right flank could be visualized from 8 h and the activity was higher than that of heart until 24 h. The most clearly visualized imaging appeared at 18 h; (2) GX1

was observed binding specifically to RMEC with no positive staining observed in control group according to Immunohistochemistry and immunofluorescence, which indicated

GX1 could target retinal neovasculature of diabetic retinopathy; (3) GX1 significantly inhibited the proliferation, micro-tube formation and RXDX-106 research buy migration of RMEC or RMEC cultured with VEGF165. Conclusion: GX1 owned the ability of specific targeting of colon cancer angiogenesis in vivo, specific binding ability and antiangiogenesis to RMEC, which indicated GX1 was to be explored for effective antiangiogenesis targeting drug to tumor and diabetic retinopathy. Key Word(s): 1. GX1 peptide; 2. tumor ; 3. diabetic retinopathy; 4. antiangiogenesis; Presenting Author: YANAN HAN Additional Authors: JIPENG YIN, KAICHUN WU Corresponding Author: YANAN HAN Affiliations: Xijing Hospital of Digestive Disease Objective: Our BMS-777607 purchase group previously got a cyclic peptide GX1 which bind selectively to endothelial cells of cancer by using a Ph. D.-C7CTM Phage display peptide library. Many previous studies in vivo and in vitro showed that, GX1 could well target

to tumor and negatively regulate angiogenesis. But its receptors are still unknown.Our aim is to screen and identify the GX1 receptors by optimizing the conditions of IP, using the immortalized human umbilical vein endothelial cells (sv-HUVEC) established by our group. Methods: 1.Special marks of endothelial cell were detected by immunofluorescence; the expression and location of GX1 receptors were detected by IF and Western MCE公司 Blot.2.The candidate proteins of GX1 receptors was obtained by using IP, sliver staining, MALDI-TOF/TOF and Bioinformatics analysis.3.The expression and location of GX1 receptors and its candidate proteins were detected and compared by WB,IF, immunohistochemistry and laser scanning confocal microscope; the recognition of candidate molecules and GX1 receptor was detected by IP,WB. Results: 1.CD31 and Factor VIII expressed on sv-HUVECs, and sv-HUVEC also expressed GX1-binding proteins, which were mainly located on cytoplasm and cell membrane. WB showed that 90-130KD proteins could bind GX1 well.2.We utilized the better conditions to enrich the GX1-binding proteins, approximately a 115KD protein band.