Antiangiogenic action of Nintedanib in prostate tumor cells and in the prostate of transgenic mice for adenocarcinoma (TRAMP) in different stages of tumor development

Author(s)

Raquel Frenedoso Silva

• Document type: Doctoral Thesis
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
• Defense date: 2018-05-25

Examining board members

Valéria Helena Alves Cagnon; Murilo Vieira Geraldo; Odair Aguiar Junior; Rejane Maira Góes; Adriana Fogagnolo Maurício

• Advisor: Valéria Helena Alves Cagnon

Abstract

Prostate cancer has been widely studied due to the large number of individuals affected by this type of injury, in addition to the complexity of the organ development, considering the important epithelial-stromal interaction as the main factor for prostatic changes. Special attention has been given to the study of FGF- and VEGF-inhibitors such as Nintedanib, a compound capable of reduce angiogenesis, prevent cell proliferation and decrease tumor volume, used in the prevention, intervention and regression of prostate cancer. Since there is no direct cure through chemical agents for advanced prostate cancer, or even effective treatment therapies, it becomes necessary to prioritize new studies in animal models that mimic the disease in men. The objective herein was to evaluate the antiproliferative action of Nintedanibe in nine tumor cell lines from different tissues, besides evaluation of proliferation and cell cycle assays, motility, invasiveness and clonogenic capacity, as well as analysis of protein profile and gene expression of human prostate tumor cells submitted to the drug. The antiproliferative action of the drug was also evaluated in xenograft mouse model for prostate cancer by inoculating PC3 cells in nude mice; subsequently, tumors were submitted to immunohistochemistry and protein profile assays. In addition, Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) were treated with Nintedanibe (10 mg/kg/day) at different stages of tumor development, i.e., malignant lesions in the initial (8 weeks), intermediate (12 weeks) and advanced (22 weeks) stages. At the end of treatment, the ventral prostate was collected for structural profile analysis and expression of proteins related to cancer development and progression such as hormonal receptors AR and ERalpha, FGF-2, FGF-7, FGFR-3, VEGF, VEGFR-1, HIF-1alpha, MMP-9 and vimentin, microvessel density (labeling for CD31), and evaluation of proliferation and apoptosis through PCNA labeling and TUNEL assay, respectively. Also, possible toxic effects of the drug in the liver, testes and epididymis were evaluated, as well as the search for metastatic foci and drug action on them. Exposure of prostatic tumor cells to Nintedanibe led to reduced cell viability and clonogenicity as well as mobile and invasive potential, besides affecting the expression of crucial molecules to the cell cycle progression, leading to a G1 phase cycling arrest. In addition, the drug decreased tumor volume by interfering with cell proliferation and microvessel density in xenograft tumor, as well as reducing levels of pro-angiogenic molecules. The results also showed that the administration of Nintedanib in TRAMP mouse at the initial and intermediate stages of the prostatic carcinoma development exerted a protective effect on cancer progression, since it delayed the neoplastic transformation in the prostatic microenvironment, reducing tumor vascularization besides immunolocalization and the levels of crucial molecules for tumorigenesis. Thus, in the present preclinical study, we conclude that antiangiogenic therapy with Nintedanibe was able to delay the neoplastic transformation in the prostatic microenvironment, reducing the necessary vascularization for tumorigenesis, preventing the tumor development to advance to more aggressive stages of the disease. Nintedanibe therapy is promising for the treatment of prostate cancer, since in it did not show liver and reproductive toxicity (AU)

• FAPESP's process: 13/26677-7 - Prostate tissue response after treatment with Nintedanibe in transgenic mice for adenocarcinoma (TRAMP) at different stages of tumor development
• Grantee: Raquel Frenedoso da Silva
• Support Opportunities: Scholarships in Brazil - Doctorate