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Thesis (Ph. D.)--Michigan State University. Dept. of Biochemistry, 2007. / Title from PDF t.p. (viewed on Apr. 16, 2009) Includes bibliographical references. Also issued in print.

<p>Angiogenesis is necessary for the growth and spread of solid tumors. In these studies angiogenesis was measured in childhood malignancies in general and in Wilms’ tumor in particular, and cutting needle biopsy (CNB) specimens were evaluated for diagnosis in childhood renal tumors. </p><p>In 33 patients with Wilms’ tumor, tumor capillaries were quantified, expression of angiogenic growth factors in tumor tissue investigated, and concentrations of angiogenic growth factors in serum measured. Reference values for angiogenic growth factors were obtained in 80 healthy adults (fibroblast growth factor 2 [FGF-2], vascular endothelial growth factor A [VEGF-A]) and 94 healthy children (angiogenin [ANG], epidermal growth factor [EGF], FGF-2, hepatocyte growth factor [HGF], tumor necrosis factor alpha [TNFA] and VEGF-A) aged 0.5-18 years. These reference values were compared with values in sera taken at diagnosis in 268 children with tumors and leukemias. CNB specimens were evaluated in 25 children with renal tumors.</p><p>A large number of capillaries was an independent prognostic factor for a poor outcome in Wilms’ tumor. Angiogenic growth factors were expressed in Wilms’ tumor tissue, and elevated concentrations of HGF and VEGF-A were found in both benign and malignant tumors. HGF was increased in leukemia, and TNFA was increased in leukemia, lymphoma and neuroblastoma. CNB, which proved to be a safe procedure, had a sensitivity of 76%. </p><p>These studies have demonstrated that quantification of capillaries is a prognostic factor in Wilms’ tumor and that HGF, TNFA and VEGF-A are frequently elevated in sera from children with cancer. Quantification of capillaries in tumor tissue and of circulating angiogenic growth factors would therefore seem to be of clinical relevance in managing children with cancer.</p>

Angiogenesis is necessary for the growth and spread of solid tumors. In these studies angiogenesis was measured in childhood malignancies in general and in Wilms’ tumor in particular, and cutting needle biopsy (CNB) specimens were evaluated for diagnosis in childhood renal tumors. In 33 patients with Wilms’ tumor, tumor capillaries were quantified, expression of angiogenic growth factors in tumor tissue investigated, and concentrations of angiogenic growth factors in serum measured. Reference values for angiogenic growth factors were obtained in 80 healthy adults (fibroblast growth factor 2 [FGF-2], vascular endothelial growth factor A [VEGF-A]) and 94 healthy children (angiogenin [ANG], epidermal growth factor [EGF], FGF-2, hepatocyte growth factor [HGF], tumor necrosis factor alpha [TNFA] and VEGF-A) aged 0.5-18 years. These reference values were compared with values in sera taken at diagnosis in 268 children with tumors and leukemias. CNB specimens were evaluated in 25 children with renal tumors. A large number of capillaries was an independent prognostic factor for a poor outcome in Wilms’ tumor. Angiogenic growth factors were expressed in Wilms’ tumor tissue, and elevated concentrations of HGF and VEGF-A were found in both benign and malignant tumors. HGF was increased in leukemia, and TNFA was increased in leukemia, lymphoma and neuroblastoma. CNB, which proved to be a safe procedure, had a sensitivity of 76%. These studies have demonstrated that quantification of capillaries is a prognostic factor in Wilms’ tumor and that HGF, TNFA and VEGF-A are frequently elevated in sera from children with cancer. Quantification of capillaries in tumor tissue and of circulating angiogenic growth factors would therefore seem to be of clinical relevance in managing children with cancer.

Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 4 uppsatser.

Aim: To explore the protective effects and involved mechanisms of two angiogenic growth factors, vascular endothelial growth factor (VEGF165) and angiopoietin-1 in cardiac myoblasts. Methods: Replication-deficient adenovirus encoding for human VEGF165 (Ad-VEGF165) or angiopoietin-1 (Ad-Ang1) were transfected into H9C2 cardiac myoblasts. Recombinant adenovirus encoding for green fluorescent protein (Ad-GFP) was used as vehicle control. Twenty-four hours later, cell apoptosis was induced by 300 μmmol of H2O2. Genomic DNA was extracted and DNA fragmentation was analyzed in 1.6% agarose gels. Phosphatidylinositol-3 kinase(PI-3 K) activity and bcl-2 expression level were investigated in H9C2 after gene transfection 24 hours later by an immol/Lunoprecipitated kinase assay and Western blot assay respectively. The effect of wartmannin, a specific inhibitor of PI-3 K, on DNA fragmentation, PI-3 K activity and bcl-2 expression was also analyzed by a pre-treatment of 30 minutes before transfection. Results: Apoptotic DNA fragmentation induced by H2O2 was significantly inhibited by the transfaction of Ad-VEGF165 and/or Ad-Ang1 but then aborted by the pretreatment of wartmannin. PI-3 K activity was significantly elevated after Ad-VEGF165 + Ad-Ang1 transfection as compared to Ad-GFP transfection group(2.60 vs 1.32, P < 0.01). Anti-apoptotic factor bcl-2 expression was upregulated in Ad-VEGF165 (2.1-fold), Ad-Ang1 (1.7-fold) and Ad-VEGF165 + Ad-Ang1 (1.7-fold) treated groups as compared to Ad-GFP transfection group. Wortmannin suppressed PI-3 K activiation induced by Ad-VEGF165 (from 1.83 to 0.69, P < 0.05). Ad-Ang1 (from 1.80 to 0.97, P = 0.07) or Ad-VEGF165a + Ad-Ang1 (from 2.60 to 0.42, P < 0.01). However, upregulation of bcl-2 induced by Ad-VEGF165 and/or Ad-Ang1 was not aborted by wortmannin pretreatment. Conclusions: VEGF165 and/or Ang1 can protect cardiac myoblasts from apoptosis induced by H2O2 throught PI-3 K and bcl-2 pathway. The anti-apoptotic function of either VEGF165 or Ang1 could be served as a now therapeutic target including their angiogenic benefits.

Aim: We aimed to clone angiopoietin-1 (Ang1) and vascular endothelial growth factor (VEGF) full-length DNAs of human origin and construct replication-deficient adenovirus encoding for either of these two genes which can be potentially served for clinical applications. Methods: VEGF165 and Ang1 full-length cDNAs of human origin were amplified by RT-PCR, verified by sequencing, cloned into a pShuttle-CMV vector, recombined with a E1 and E3 regions double-deleted adenovirus, packaged in 293A cells, and purified by ultracentrifugation. The titers of Ad-Ang1 and Ad-VEGF165 were determined by a tissue culture infectious dose50 method. Expression of Ang1 and VEGF165 proteins in H9C2 cardiac myoblasts was examined by Western blot. To examine the protective properties of Ad-Ang1 and Ad-VEGF165, DNA fragmentation induced by H2O2 was analyzed in H9C2 cells 24 hours after transfection. Ad-GFP served as a vehicle control. Results: Sequencing analysis indicated that there is one base difference at site 1206 (t) in Ang1 compared with that of GeneBank (c, U83508) although the coded amino acids are the same (Ileucine). VEGF165 cDNA sequence was same as that of GeneBank (AB021221). Western blot showed that protein levels of Ang1 and VEGF165 were increased 3.53 and 11.53 fold respectively 24 h after transfection as compared to control. Examination of DNA fragmentation suggested that Ang1 and/or VEGF165 significantly protected H9C2 cells from H2O2 induced apoptosis. Conclusions: The two constructed adenoviral vectors, Ad-Ang1 and Ad-VEGF165, functionally expressed target proteins. We demonstrated, for the first time, that the combined utilization of Ang1 and VEGF165 inhibited apoptosis, in addition to their angiogenesis properties.