Expression Of Angiopoietin-1, 2,VEGF In Larynx Squamous Cell Carcinoma And Their Correlations With Angiogenesis

Background and objectiveLaryngeal cancer is one of the common malignancy tumors in the otolaryngology head and neck surgery,mainly is Larynx Squamous Cell Carcinoma. Although in recent years the effective of Laryngeal cancer treatment and laryngeal function of retention rate has improved because the therapy methods increased, but the five-year survival rate had not increased significantly. Especially the patients with cervical lymph node metastasis and distant metastasis had the more worse prognosis. Therefore clinical studies focus on the incidence of Laryngeal cancer, the development of Laryngeal cancer and transfer of law and gene therapy. With the progress of molecular biology, people growingly realized that oncogenes and the expression of promoting angiogenesis factor were closely related to the incidence of tumors and the development of the tumor.There may be the synergistic effect between them. The present study focused on the mechanism of angiogenesis in vascular endothelial growth factor (vascular endothelial growth factor, VEGF) and its receptors. Another family of systems which was recently discovered and was associated with angiogenesis, vascular endothelial growth factor (also called angiogenesis factors. angiopoietin) and its receptor system,has to be focused.Ang family is a newly discovered protein family, only with vascular endothelial growth factor receptor agonist and antagonist. The Ang family include angiopoietin-1, 2, 3, 4.We major study on the angiopoietin-1,2 which mainly express in embryonic development and promote the development and maturing of the cardiovascular system.In addition to the higher expression in adult female reproductive system, other organizations showed low expression. Angs play an important role in the maintenance of vascular integrity mature in physiological, involved in the menstrual cycle and physical rehabilitation of endometrial angiogenesis. Reaserches discovered that Angs have played an important role in tumor angiogenesis in recent years. Ang family with the process of interaction with VEGF in tumor angiogenesis promotes the formation of angiogenesis. Newly discovered angiopoietin-2 (Ang-2) is a secreted protein. Ang-2 affect tumor growth,metastasis and angiogenesis in the tumor angiogenesis by strengthening the role that VEGF induced angiogenesis.We have not found any research about the relationship between the expression of Ang with the expression of VEGF and any research about the relationship between the expression of Ang and VEGF with angiogenesis in LSCC.At present,IMVD has been a golden standard which evaluates tumor angiogenesis. But there is controversy that Squamous cell carcinoma of the head and neck tumor microvessel density(intratumor microvessel density, IMVD) is related to the prognosis. This may be due to vascular endothelial cell markers used in different ways and different organizations pretreatment and Microvessel counting or non-standard methods. Von Willebrand factor, CD31 and CD34 are the markers of all vessel endothelial cells. CD105 is different from above.Its antibody firstly combine activated endothelial cells and the formation of vascular endothelial cells. To further investigate the relationship between IMVD with the prognosis of head and neck squamous cell carcinoma, We detect the 57 cases of LSCC with CD105 antibody through Immunohistochemical method and count IMVD and analysis theirs relationships.Materials and MedthodsDetecting the expression of Ang-1,2,VEGF and the microvascular density markered by CD105 through Immunohistochemical method in 57 cases of LSCC specimens,7 cases normal tissues surrounding the LSCC and 9 cases vocal polyps and analysising the relationship between the pathological features of laryngeal carcinoma with theirs expression through Statistical Methods..Results1. Ang-1,2 expressed in LSCC and normal epithelial cells and vascular endothelialcells, mainly localized in the cytoplasm.2.The expression Ang-1,2 in laryngeal squamous cell carcinoma were 68.85% and62.30% respectively, significantly higher than the normal tissues surroundingLSCC,vocal polyps (P<0.05),but there was no difference between the normal tissuessurrounding LSCC and vocal polyps(P>0.05).3.CD105 expressed in all the specimens, CD105-positive staining (brown) mainlylocated in the vascular endothelial cell membrane and the cytoplasm.The IMVD ofthe surrounding of cancer nests was higher than the IMVD of the central of cancernests,but the endothelial cell membrane and the cytoplasm of the mature vascularwere not expressed.4. The IMVD marked by CD105 in the period of T3-T4 were significantly higher than that in the period of T1-T2 (P<0.05). The IMVD marked by CD105 was increased with the increase of clinical stages (P<0.05).There was no significant difference between the IMVD of cervical lymph node metastasis with the IMVD without lymph node metastasis (P<0.05). The IMVD of the group of moderiatly and poorly differentiated was higher than the IMVD of the group of the well differentiated,but the difference was not statistically significant (P>0.05).5. VEGF expressed in LSCC and vocal polyps epithelial cells and the normal tissues surrounding the LSCC, mainly localized in the cytoplasm and nuclear.The positive rate of VEGF was 73.68% (42/57) in LSCC.The positive rate of VEGF in the adjacent normal tissue and vocal polyps were 28.57% and 33.33%. The positive rate LSCC was higher than that in the adjacent normal tissue and vocal polyps, the difference was significant (P<0.05),but there was no difference between the adjacent normal tissue with vocal polyps(P>0.05).The expression of VEGF in LSCC was related to cervical lymph node metastasis,histological grades,clinical stages and T stages, the differences was statistically significant (P<0.05).6. The expression of Ang-2 was related to cervical lymph node metastasis,clinical stages and T stages, the differences has statistically significant (P<0.05),but the expression of Ang-1 was only reated to the clinical stages. The expression of Ang-1,2 had no differences in histological grades and aged stages(.P>0.05).7. The IMVD with Ang-2-positive and the IMVD with VEGF-positive in LSCC were significantly higher than the IMVD with Ang-2-negative and the IMVD with VEGF-negative in LSCC (6.7053±0.6900 VS 5.9900±1.3386, 6.7379±0.6805 VS 5.7153±1.4142, P=0.031,0.0160.05). There was no significant difference between The IMVD with Ang-1 positive in LSCC with the IMVD with Ang-1- negative in LSCC (6.7106±0.8343 VS 6.4195±0.9938, P=0.286>0.05) .8. The IMVD with both of Ang-1 -positive and Ang-1 -positive (6.6175±0.4832) was significantly higher than the IMVD with Ang-1-positive and Ang-2-negative(4.8627±0.5595) and the IMVD with both of Ang-1-negative and Ang-1-negative (6.0390±0.2465, P=0.000, 0.003 <0.05 ) . But there was no significantly betweenthat with the IMVD with Ang-2-positive and Ang-1-negative ( 6.8938±0.6626,P=0.171>0.05 ) .9. The IMVD with Ang-2-positive and VEGF-positive was the maximum (6. 7748±0.7184) and significantly higher than any other states (P=0.008, 0.000, 0.000 <0.05).Conclusions1.The expression of Ang-1,2 in LSCC was significantly higher than that in othergrounps.These suggested that Ang-1 and Ang-2 may take participate in the occurrenceand development of LSCC.2.The expression of Ang-2 in LSCC was related to the clinical stages,the scope ofthe invasion increasing and lymph node metastasis. Ang-2 may promote the invasionand the metastasis of LSCC and can be used as one of the predictors of prognosis inLSCC. 3.The IMVD marked with CD105 has significant correlation with clinical stage and the scope of the invasion increasing. CD105 also can be used as one of the predictors of prognosis in LSCC.4. The expression of Ang-2 and VEGF were significantly correlated with the IMVD.The IMVD of Ang-2-positive and VEGF-positive significantly higher than the IMVD of Ang-2 negative and VEGF-negative. Ang-2 and VEGF may promote angiogenesis in LSCC.5. The IMVD of Ang-2-positive and VEGF-positive was the highest in all the states.That indicated that the expression of Ang-2 and VEGF may have a synergistic effect which can promote angiogenesis.6. Ang-2 and VEGF may affect the invasion and the metastasis of LSCC through promoting angiogenesis.