Inhibition of the Tcf/beta-catenin complex increases apoptosis and impairs adrenocortical tumor cell proliferation and adrenal steroidogenesis

Author(s)

Leal, Leticia F. ^[1] ; Bueno, Ana Carolina ^[1] ; Gomes, Debora C. ^{[1, 2]} ; Abduch, Rafael ^[1] ; de Castro, Margaret ^[3] ; Antonini, Sonir R. ^[1]

Affiliation

^[1] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Pediat, Sao Paulo - Brazil

^[2] Univ Fed Uberlandia, Sch Med, Dept Pediat, BR-38400 Uberlandia, MG - Brazil

^[3] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Internal Med, Sao Paulo - Brazil

• Document type: Journal article
• Source: ONCOTARGET; v. 6, n. 40, p. 43016-43032, DEC 15 2015.
• Web of Science Citations: 24

Abstract

Background: To date, there is no effective therapy for patients with advanced/metastatic adrenocortical cancer (ACC). The activation of the Wnt/beta-catenin signaling is frequent in ACC and this pathway is a promising therapeutic target. Aim: To investigate the effects of the inhibition of the Wnt/beta-catenin in ACC cells. Methods: Adrenal (NCI-H295 and Y1) and non-adrenal (HeLa) cell lines were treated with PNU-74654 (5-200 mu M) for 24-96 h to assess cell viability (MTS-based assay), apoptosis (Annexin V), expression/localization of beta-catenin (qPCR, immunofluorescence, immunocytochemistry and western blot), expression of beta-catenin target genes (qPCR and western blot), and adrenal steroidogenesis (radioimmunoassay, qPCR and western blot). Results: In NCI-H295 cells, PNU-74654 significantly decreased cell proliferation 96 h after treatment, increased early and late apoptosis, decreased nuclear beta-catenin accumulation, impaired CTNNB1/beta-catenin expression and increased beta-catenin target genes 48 h after treatment. No effects were observed on HeLa cells. In NCI-H295 cells, PNU-74654 decreased cortisol, testosterone and androstenedione secretion 24 and 48 h after treatment. Additionally, in NCI-H295 cells, PNU-74654 decreased SF1 and CYP21A2 mRNA expression as well as the protein levels of STAR and aldosterone synthase 48 h after treatment. In Y1 cells, PNU-74654 impaired corticosterone secretion 24 h after treatment but did not decrease cell viability. Conclusions: Blocking the Tcf/beta-catenin complex inhibits the Wnt/-beta-catenin signaling in adrenocortical tumor cells triggering increased apoptosis, decreased cell viability and impairment of adrenal steroidogenesis. These promising findings pave the way for further experiments inhibiting the Wnt/beta-catenin pathway in pre-clinical models of ACC. The inhibition of this pathway may become a promising adjuvant therapy for patients with ACC. (AU)

• FAPESP's process: 11/13807-4 - Analysis of the WNT and Sonic Hedgehog pathways in childhood adrenocortical tumors and its relationship with miRNAs
• Grantee: Sonir Roberto Rauber Antonini
• Support Opportunities: Regular Research Grants
• FAPESP's process: 11/10512-3 - Analysis of Wnt/{beta}-catenin pathway in childhood adrencortical tumors
• Grantee: Letícia Ferro Leal
• Support Opportunities: Scholarships in Brazil - Doctorate