The Role Of RhoC In Invasiveness Of Venous Malformation In Soft Tissue

PART ONE The Expression of RhoC in Venous malformation Tissue Objective To indentify the character of venous malformation pathology and investigate the expression of RhoC in venous malformation tissues and its significance.Methods Specimens of15invasive venous malformation tissues,12non-invasive venous malformation tissues and12normal subcutaneous tissues were collected and stained with H.E in order to observe the pathological change of the tissue morphology. The protein expression of RhoC was detected by Western Blot while the expression of RhoC mRNA was detected by PCR.Results Observed under microscope, venous malformations were mainly formed by irregular expansion, derangement vessels and connective tissue, which scattered with adipose tissue. The vascular was abnormal shaped, with large cavity and thin wall, the endothelial cells arranged disorderly. The RhoC protein and mRNA were expressed in all three tissues. In the invasive venous malformation tissues, the protein and mRNA levels were significantly higher than the others(P<0.05).Conclusion The express of RhoC in the invasive venous malformation tissues is higher. RhoC may be involved in the invasive development of the venous malformation. PART TWO The Effect of RhoC on Invasion Behavior of Venous Malformations Endothelial CellsObjective To investigate the change of invasion ability and tube formation capacity after transfected RhoC-siRNA to VM-EC.Methods After Venous malformation endothelial cells(VM-EC) and human umbilical vein endothelial cells(HUVEC) were isolated and cultured, the protein and mRNA expression of RhoC in cells were detected. Transfect RhoC-siRNA to VM-EC and inhibit the expression of RhoC, VM-EC were cultured to investigate the ability of invasion and tube formation by Invasion Assay and Network Formation Assay.Results The Primary cultured VM-EC and HUVEC were Short spindle, both IMVM-ECs and HUVECs displayed the positive staining of FⅧ-Ag. The expression of RhoC in VM-EC was significantly higher than in HUVEC(P<0.05). After transfected RhoC-siRNA to VM-EC, the expression of RhoC is decreased significantly. In the Invasion Assay, the RhoCRNAi+cells which penetrated the Matrigel gelatin and filterable membrane, and arrived the under membrane were less than RhoCRNAi-cells; in Network Formation Assay, the integrity Vascular ring formed by RhoCRNAi+cells were less than RhoCRNAi-cells.Conclusion The decreased expression of RhoC may attenuate the invasive behavior of the VM-EC. PART THREE The Impact of RhoC on MMP-9in Venous Malformation endothelial cellsObjective To investigate the expression of MMP-9in VM-EC and observe the impact of RhoC on the expression of MMP-9.Methods Culture VM-EC and HUVEC, the protein expression of MMP-9was detected by Western Blot while the expression of RhoC mRNA was detected by PCR. Transfect RhoC-siRNA to VM-EC, detect the protein and mRNA expression of MMP-9in VM-EC.Results The expression of MMP-9in VM-EC was significantly higher than in HUVEC(P<0.05). After transfected RhoC-siRNA to VM-EC, the expression of RhoC and MMP-9were significantly lower than untransfected group (P<0.05).Conclusion RhoC inhibited can decline the expression of MMP-9, MMP-9maybe one of ways which RhoC influence the invasive progress of venous malformation.