RaplGAP Inhibited The Proliferation,Migration And Invasion Abilities Of HeLa

Objective: To explore the effects of Rap1GAP on the proliferation, migration and invasion abilities of HeLa and the molecular mechanism of inhibiting invasion. Cervical cancer ranks second only to breast cancer in women malignant tumors globally. In recent years, the incidence increases significantly with increasing incidence rate, earlier age of onset and aging. To inhibit uterine cervix cancer cell proliferation, invasion and metastasis has the vital significance for malignant tumor treatment and enhances cancer patients'survival rate. Tumor genesis is regulated by gene abnormality in cell proliferation, apoptosis, gene stability, angiogenesis, invasion or metastasis. Anti-oncogene inhibits tumor metastasis during the multi-factor and multi-stage process. Rap1GAP ( Rap1 GTPase-activating protein 1) is a family of proteins with GAP function specific for Rap1,which catalyzes activated Rap1-GTP into inactinated Rap1-GDP. Rap1 is a small GTP-binding protein, which controls proliferation, differentiation and adhesion-related functions such as cell-cell contacts and functional activation of integrins. Disturbance of active Rap1 contributes to the pathogenesis and development of malignant tumors, therefore, as a regulator during Rap1 activation, Rap1GAP will prove to be one highly relevant factor in the same process.Effects of Rap1 are cell type specific. For example, overexpression of Rap1 in Swiss 3T3 cells induces DNA synthesis and decreases doubling time, but inhibits proliferation in Ras transfected NIH3T3 cells and astrocytes. Accordingly, the function of Rap1GAP is probably cell type specific. This article explores the effects of Rap1GAP on the proliferation,migration and invasion abilities of HeLa and the molecular mechanism of inhibiting invasion.Methods: The uterine cervix cancer HeLa cells, HeLa cells stably expressing GFP and HeLa cells stably expressing Rap1GAP were cultured in vitro. The effects of Rap1GAP on proliferation in HeLa were detected by MTT test. To study the effects of the Rap1GAP on inhibiting HeLa adhere, invade and migrate through cell adhersion assay, cell wound healing assay , matrigel invasion assay and transwell migration assay. MMP-2 and MMP-9 was detected by gelatin zymography.Results:1.Compared with control groups(untransfected group and empty plasmid transfected group), growth velocity decreased significantly( p<0.05) . 2.The adherent rate to FN in HeLa-Rap1GAP decreased compared with control groups (p<0.05). 3.There was statistical difference between HeLa-Rap1GAP and control groups in Migration distance. 4.Cells counts were much less in HeLa-Rap1GAP than in control groups in matrigel invasion assay and transwell migration assay. 5.The synthesis and activation of MMP-2 and MMP-9 in HeLa-Rap1GAP was lower.Conclusions:1.Rap1GAP can obviously inhibit the proliferation of human uterine cervix cancer cell line HeLa in vitro.2.Rap1GAP inhibited the migration of uterine cervix cancer cell line HeLa in vitro.3.Rap1GAP inhibited the invasion of uterine cervix cancer cell line HeLa in vitro.4.Suppressing the expression of MMP-2 and MMP-9 may be a feasible way, by which Rap1GAP educes its inhibiting effect in HeLa cells invasion.