Influence Of Preoperative Chemotherapy On VEGF And Ang2 Expression And Angiogenesis Of Gestational Trophoblast Tumor

ObjectiveIt is well known that tumor growth and metastasis depends on tumor angiogenesis . A lot of evidence exists to show that there is a consistently strong relation between microvessel intensity and tumor growth and metastasis. There will be a lot of angiogenic factors secreted in tumor growth . But factors specifically acted on endothelial cells including vascular endothelial growth (VEGF), angiopoitins and Ephrin can enhance angiogenesis via binding with their receptors in endothelial cells. There is very important binding VEGF with its receptors in angiogenesis. But maturation and stabilization of vessel depends on maintenance of angiopoietin (Angl,Ang2) and their receptor(Tie2). Their expression elevated in many malignant tumors and were considered to relate to tumor growth and angiogenesis.There are a lot of reports about microvessel density and angiogenesis cytokine in kinds of solide tumors, however few in gestational trophoblast neoplasm(GTN). Gestational trophoblast neoplasm is different from the other solid tumors in its angiotrophic, invading vases and taking place of endothelium cells to form vascular wall, affluent vases in the focus. Preoperative chemotherapy makes a lot of tumor cells necrosis, vases decrease and micro-thrombus of capillary vessel form to ensure the operation go on wheels, hemorrhage reduce and simultaneously restrain tumor growth,metastasis. This study discusses the influence of preoperative chemotherapy on VEGF and Ang2 expression and angiogenesis of gestational trophoblast neoplasm by immunohistochemistry, perhaps provides theoretic gist for preoperative chemotherapy. Methods1. Object: 74 cases of gestational trophoblast neoplasm. Forty-seven of these performing preoperative chemotherapy in Women's hospital, School of Medicine, Zhejiang University, from 1996.1 to 2002.12.(23 cases of choriocarcinoma, 24 cases of invasive mole) were regarded as chemotherapy group, preoperatively given EMA-Co or ACM project for 2~3 periods of treatment. During the corresponding period, the other 27 cases hysterectomized but not performed chemotherapy were regarded ascontrol group (5 cases of choriocarcinoma and 5 cases of invasive mole from Second affiliated hospital , School of Medicin, Zhejiang University). Samples were fixed in formalin, embedded in paraffin, sliced up routinely, HE staining and evidenced by pathology. All of the pathologic slices were checked by repeats. The average age of chemotherapy group was 33 6 years old, including 14 cases of clinical grade 1,12 cases of grade II, 17 cases of grade III, and 4 cases of grade IV. WHO prognosis grade: 16 cases of high risk, 15 cases of middle risk, and 16 cases of low risk. The average age of control group was 34 + 7 years old, including 8 cases of clinical grade I , 5 cases of grade II, 12 cases of grade III, and 2 cases of grade IV. WHO prognosis grade: 12 cases of high risk, 7 cases of middle risk, and 8 cases of low risk. Both groups did not perform other assistant treats. There is no statistical difference between patients' clinical pathologic factors of the two groups.2. Pathological examinationParaffin slices about 5m thickness were undergone dimethylbenzene dewaxed, grads ethanol dehydrated, HE dyed, grads ethanol clarified and sealed. Slices were observed by optical microscope.3. ImmunohistochemistryParaffin slices fixed in formaldehyde were sliced up continuously, about 4 - 6m, undergone dimethylbenzene dewaxed, grads ethanoldehydrated, and antigen restored. Clear up endogenetic peroxidase activation with 3% hydrogen peroxide for 5min; apply 7.5% goat serum to block nonspecific secondary antibody-binding sites for 30min at 37; apply primary antibody of different dilute degree; incubate for 30min at 37 (polyclonal antiboby) or 4 overnight (monoclonal antibody), and manipulate according to the introduction. Count percent grade of VEGF and Ang2 stained tumor cells under 400X field by semi-quantify method and select five fields of 200X to count CD34(+) cells.4. Statistics method:Metri...