Study On The Role Of Vegf-c,angiogenin-1 And Endostatin In The Formation Of Lymphatic Malformation Of Oromaxillo-facial Region

BackgroundLymphatic malformation of oromaxillo-facial region often results to 'huge tongue','huge lip' and brings great pressure and psychological burden to the patients and their families.With checking into the disease,a viewpoint has been received by more and more scholars.The view asserts that the formation of lymphatic malformation dues to the break of the balance of which is regulated and controlled by the promoting factors and inhibiting factors of the formation by genetic mutation. After analysising the current situation of the study of the disease and according to the view that genetic mutation leads to the change of the number of relevant endothelial cells,our study plans to explore the expression of relevant factors of the formation of lymphatic vessels and blood vessels in the formation of lymphatic malformation of oromaxillo-facial region mainly and then explore the causes of vascular malformation.VEGF-C(vascular endothelial growth factor-C) is one of the most important growth factor of lymphatic vessel so far.VEGF-C which is secreted by tumor cells can integrate VEGFR-3(vascular endothelial growth factor receptor-3) and then mediate lymphatic vessels' formation by promote LEC(lymphatic endothelial cell) 's proliferation,migration and survivorship.VEGF-C with complete structure integrates VEGFR-3 only,while mature segments of VEGF-C after processing have remarkably increasing bonding force with VEGFR-3.Homozygous deletion of VEGF-C can lead to deficiency of formation of lymphatic vessels of mice,which points out that VEGF-C is necessary in the process of lymphatic vessels' growth of embryoes of mice.It is also found by researches that VEGF-C can start the signal of formation of lymphatic vessels by integrates other receptors.Furthermore,Sidle DM discovered that strong expression of VEGF-C is related to the recurrence of lymphatic malformation.Angiogenin(Angiogenin,Angiopoietin,ANG) is a family of protein found newly and it is a type of growth factors srcreted by tumor cells.It can promote the growth of neonatal blood vessels and including four molecules:ANG-1,ANG-2,ANG-3 and ANG-4.Ang acts on Tie-2(tie-2 receptor) which expressions in the endothelial cells specially to keep the completeness and resting situation of mature blood vessels and participates the newborn of blood vessels in physiological and pathological situations.In the view of current researches,there is a bad relationship between the expression of ANG-1 and the formation of blood vessels of tumor but the expression of ANG-2 is a factor of the beginning and strengthening of the newborn of blood vessels of tumor and is closely related to the number of neonatal blood vessels,clinical stages and prognosis of tumor.It has been approved that ANG-1 can promote the growth of LEC and the formation of lymphatic vessels in the esoteric examinations and exoteric ones.Endostatin(END) was seperated from the blood serum of mice which suffer EOMA by O'Reilly in 1997 for the first time.It is the degradate product of collagen X (nearⅦ)and can be the inhibition of the formation of blood vessels which roots in tumor.It is found that END can inhibite the proliferation and migration of LEC intensively in vitro by researches.It can be beheld soon that the death of LEC after little END is added in and the number of dead cells increases obviously with the concentration of END rises.It might be similar to the mechanism of the action which END acts on blood vascular endothelial cells.The following is the process of the latter: END integrates the receptors in the surface of END cells and thus inhibites the hyperplasia of endothelial cells;END prevents bFGF from integrating its receptors so that block the access of the signal transduction;END can inhibite the IAPs as Bcl-2 and Bcl-x1 and then accelerate the apoptosis of endothelial cells;END can activate FAK directly and wreck the formation of dots of endothelial cells,so that it inhibite the migration of endothelial cells and at last the cells turn into round,leave the wall,float and die step by step.ObjectiveTo explore the expression of VEGF-C,ANG-1 and END in the lymphatic malformation of oromaxillo-facial region and the relation between their expression and the occurence,growth and prognosis of the disease.The experiment selected organizations of lymphatic malformation from different category and positions and normal organizations as objects.We detected the expression of the three proteins by immunochemistry(SABC) separately in order to find our their change and interrelation during the disease's occurence and growth.Meterials and methods1.61 specimens of lymphytic malformation of oromaxillo-facial region have been gathered from the Department of Pathology,the Fist Affiliated Hospital of Zhengzhou University from July 2002 to December 2006.The specimens treated by mineral wax were:14 cases of microcapsule,30 cases of macrocapsule,17 cases of lymphartic malformation and venule malformation.They were 29 cases of tongue,5 cases of the parotid region,18 cases of the submental region and upper hind neck,9 cases of cheek. The patients are 2 months to 66 years old and their median is 13.9 years old.They were all not treated by sclerotherapy or others treatments.10 normal specimens of oromaxillo-facial region were the group of negtive control.They were all fixed by neutral formalin solution(PH=7.4),evaporated routinely and then were paraffined imbedded.2.To detect the expression of VEGF-C,ANG-1 and END separately by immunochemistry(SA1022). 3.The staining results were analysed by computer with the Quantitative Analysis System in computer.The results were analysed by SPSS.14.0 software package. Student's t-test,rank sum test,chi-square test and Fisher's exact test of probabilities were used for data analysis.Possible correlations between variables of the analyzed samples were examined by Spearmans test.Statistically significant level was considered as "alpha equals 0.05".Results1.The positive staining of VEGF-C was mainly observed in the cytoplasm and a few in the epicyte.The staining appears buffy,granular and dispersed.Expression of VEGF-C in the normal specimens was weak.There were significant differences between lymphatic malformation and normal tissues(p＜0.05).There were no significant differences between macrocapsule and microcapsule(p＞0.05).But there were significant differences between either of them and normal tissues(p＜0.05).2.The positive staining of ANG-1 was mainly observed in the cytoplasma and a few in the epicyte.The staining appears buffy,granular and dispersed.Expression of ANG-1 in the normal specimens was weak.There were significant differences between lymphatic malformation and normal tissues(p＜0.05).There were no significant differences between macrocapsule and microcapsule(p＞0.05).But there were significant differences between either of them and normal tissues(p＜0.05).3.The positive staining of END was mainly observed in the cytoplasm and a few in the epicyte.The staining appears buffy,granular and dispersed.Expression of END in the normal specimens was not weak while expression in lymphatic malformation was weak.There were significant differences between lymphatic malformation and normal tissues(p＜0.05).There were no significant differences between macrocapsule and microcapsule(p＞0.05).But there were significant differences between either of them and normal tissues(p＜0.05).4.Spearmans test shows that there were interrelation between the expression of VEGF-C and ANG-1;VEGF-C and END in lymphatic malformations. Conclusions1.There is a certain relationship between the expression of VEGF-C and the disease of lymphatic malformation.The abnormal expression of VEGF-C maybe act in the course of the occurence and growth of lymphatic malformation.VEGF-C maybe one of the index of the research of etiology of lymphatic malformation.2.There is a certain relationship between the expression of ANG-1 and the disease of lymphatic malformation.The abnormal expression of ANG-1 maybe act in the course of the occurence and growth of lymphatic malformation.ANG-1 maybe one of the index of the research of etiology of lymphatic malformation.3.There is a certain relationship between the expression of END and the disease of lymphatic malformation.The abnormal expression of END maybe act in the course of the occurence and growth of lymphatic malformation.END maybe one of the index of the research of etiology of lymphatic malformation.4.There were interrelation between the expression of VEGF-C and ANG-1; VEGF-C and END in lymphatic malformations.5.The combined detecting of the expression of VEGF-C,ANG-1 and END by immunochemistry staining together may contributes to the filtration,treatment and estimation of prognosis of the disease of lymphatic malformation.