The Inhibitory Effect And Mechanism Of RLj-RGD3 On Ovarian Cancer SKOV3 Cells And The Nude-mouse Transplanted Tumor Model

rLj-RGD3 is a recombinant protein with the molecular weight of 13.5 k Da,which contains 3 RGD motifs and identified from the buccal gland of Japanese lamprey.Previous studies have shown that rLj-RGD3 has the broad-spectrum anti-tumor activities,as well as the anti-angiogenesis property.However,there were not studies and reports about the effect of rLj-RGD3 on human ovarian cancer SKOV3 cells.In order to study the pharmacology and pharmacodynamics of rLj-RGD3 on human ovarian cancer SKOV3 cells,the inhibitory effects and mechanisms of rLj-RGD3 on human ovarian cancer SKOV3 cells and the transplanted tumor of nude mice bearing SKOV3 were studied in the present study.rLj-RGD3 was purified as a single band on SDS-PAGE through affinity chromatography due to its high concent of histidines,which provided the information for its application.Cell proliferation assay in vitro showed that rLj-RGD3 could inhibit SKOV3 proliferation in a dose dependent and IC₅₀value of rLj-RGD3 in SKOV3 cells was9.7μM.In order to detect whether rLj-RGD3 could induce the apoptosis of SKOV3 cells,TUNEL,Hochest33258 staining,flow cytometry（Annexin V/PI）and western Blot were used.And the results of those experiments showed that rLj-RGD3 induced apoptosis of SKOV3cells through multiple signaling pathways,including the mitochondrial apoptosis pathway,the death receptor pathway and the MAPK signaling pathways.In addition,cell wound scratch,transwell assays,phalloidine labeling and western blot showed that rLj-RGD3 inhibited basic fibroblast growth factor（b FGF）-induced migration and invasion of SKOV3 cells in a dose-dependent manner by destroying the cytoskeleton,supressing the pseudopod formation and reducing the expression and phosphorlation levels of focal adhesion kinase（FAK）in SKOV3 cells.On the basis of above assays in vitro,the transplanted tumor model of nude-mice with SKOV3 was established in the present study.The results of the in vivo assays showed that rLj-RGD3 suppressed the tumor growth effectively without affecting the weight of nude-mouse.In conclusion,rLj-RGD3 could inhibit the activities and functions of human ovarian cancer SKOV3,which would become a potential candidate drug of anti-tumor.