Studies on resistance to apoptosis in cancer, head/neck model: signaling via with emphasis on PIP3-Akt/SET, oxidative stress, mitochondria and relationships

• Grant number: 09/52228-0
• Support type: Research Projects - Thematic Grants
• Duration: March 01, 2010 - May 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Metabolism and Bioenergetics
• Principal researcher: Carlos Curti
• Grantee: Carlos Curti
• Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Tumor cells, in particular of head and neck scamous cell carcinoma (HNSCC), present as important characteristic the resistance to undergo apoptosis, which reflects both in aggressiveness of cancer and inefficiency of treatment against the disease. The present study proposes to assess molecular mechanisms of resistance of apoptosis signaling in HN13 cells (oral scamous cell carcinoma) compared to HEK293T cells (renal embryonic, as non tumoral control), by means of the process induced by tert-butyl hydroperoxide and stauroporin. Biochemical and molecular methods, involving approach in cancer, mitochondria, oxidative stress and cell death, will be employed in the following assessments: i) Akt activation via phosphorylation and its inactivation by PP2A via desphosphorylation inhibited by SET; ii) confluence of Akt phosphorylation to the mitochondrial mechanisms that modulates apoptosis; iii) involvement of reactive oxygen species (ROS); iv) identification of strategic regulation targets of signaling pathways involving oxidative stress/antioxidant defense, mitochondria, apoptosis, and cancer pathway finder, to be defined on account of preliminary analyses by PCR Arrays Real time systems associated with RNAi/knockdown technique and ectopic expression of protein. The apoptosis studies include cell viability, phosphatidil-serine exposition, DNA fragmentation, caspase 9, 8 and 3 activation, as well as generation/accumulation of ROS, oxidized/reduced glutatione levels and superoxide dismutase activity. The mitochondrial studies include expression of the proteins Bax and Bcl-2 oligomerization of Bak with Bax or with Bcl-2/Bcl-XL and their interactions with mitochondria, release of apoptogenic factors from the organelle (citochrome c, AIF), mitochondrial membrane potential and permeability transition, ATP levels and interaction of hexoquinase II with mitochondria. (AU)

Scientific publications

(8)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ALMEIDA, LUCIANA O.; NETO, MARINALDO P. C.; SOUSA, LUCAS O.; TANNOUS, MARYNA A.; CURTI, CARLOS; LEOPOLDINO, ANDREIA M.. SET oncoprotein accumulation regulates transcription through DNA demethylation and histone hypoacetylation. ONCOTARGET, v. 8, n. 16, p. 26802-26818, APR 18 2017. (13/10898-4, 10/20536-4, 13/08135-2, 10/20384-0, 09/52228-0, 09/10783-7)

SOBRAL, LAYS M.; SOUSA, LUCAS O.; COLETTA, RICARDO D.; CABRAL, HAMILTON; GREENE, LEWIS J.; TAJARA, ELOIZA H.; GUTKIND, J. SILVIO; CURTI, CARLOS; LEOPOLDINO, ANDREIA M.. Stable SET knockdown in head and neck squamous cell carcinoma promotes cell invasion and the mesenchymal-like phenotype in vitro, as well as necrosis, cisplatin sensitivity and lymph node metastasis in xenograft tumor models. Molecular Cancer, v. 13, FEB 20 2014. (10/18544-9, 13/10898-4, 09/52228-0, 10/20384-0, 13/08135-2)

ALMEIDA, LUCIANA O.; GARCIA, CRISTIANA B.; MATOS-SILVA, FLAVIA A.; CURTI, CARLOS; LEOPOLDINO, ANDREIA M.. Accumulated SET protein up-regulates and interacts with hnRNPK, increasing its binding to nucleic acids, the Bcl-xS repression, and cellular proliferation. Biochemical and Biophysical Research Communications, v. 445, n. 1, p. 196-202, FEB 28 2014. (07/02223-6, 13/10898-4, 09/52228-0, 10/20384-0, 09/10783-7)

LEOPOLDINO, ANDREIA M.; SQUARIZE, CRISTIANE H.; GARCIA, CRISTIANA B.; ALMEIDA, LUCIANA O.; PESTANA, CEZAR R.; SOBRAL, LAYS M.; UYEMURA, SERGIO A.; TAJARA, ELOIZA H.; GUTKIND, J. SILVIO; CURTI, CARLOS. SET protein accumulates in HNSCC and contributes to cell survival: Antioxidant defense, Akt phosphorylation and AVOs acidification. Oral Oncology, v. 48, n. 11, p. 1106-1113, NOV 2012. (05/03380-2, 10/20384-0, 06/06334-4, 09/52228-0)

ALMEIDA, LUCIANA O.; GOTO, RENATA N.; NETO, MARINALDO P. C.; SOUSA, LUCAS O.; CURTI, CARLOS; LEOPOLDINO, ANDREIA M.. SET overexpression in HEK293 cells regulates mitochondrial uncoupling proteins levels within a mitochondrial fission/reduced autophagic flux scenario. Biochemical and Biophysical Research Communications, v. 458, n. 2, p. 300-306, MAR 6 2015. (13/01355-7, 13/10898-4, 09/52228-0, 10/20384-0, 09/10783-7)

OUCHIDA, AMANDA TOMIE; UYEMURA, VALERIA TUDELLA; QUEIROZ, ANDRE LIMA; BRAUER, VERONICA SOARES; CAVALCANTI-NETO, MARINALDO PACIFICO; SOUSA, LUCAS OLIVEIRA; UYEMURA, SERGIO AKIRA; CURTI, CARLOS; LEOPOLDINO, ANDREIA MACHADO. SET protein accumulation prevents cell death in head and neck squamous cell carcinoma through regulation of redox state and autophagy. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, v. 1866, n. 4, p. 623-637, APR 2019. (13/10898-4, 16/19103-2, 09/52228-0)

LEOPOLDINO, ANDREIA M.; SQUARIZE, CRISTIANE H.; GARCIA, CRISTIANA B.; ALMEIDA, LUCIANA O.; PESTANA, CEZAR R.; POLIZELLO, ANA C. M.; UYEMURA, SERGIO A.; TAJARA, ELOIZA H.; GUTKIND, J. SILVIO; CURTI, CARLOS. Accumulation of the SET protein in HEK293T cells and mild oxidative stress: cell survival or death signaling. Molecular and Cellular Biochemistry, v. 363, n. 1-2, p. 65-74, APR 2012. (05/03380-2, 06/06334-4, 09/52228-0)

ALMEIDA, LUCIANA O.; GOTO, RENATA N.; PESTANA, CEZAR R.; UYEMURA, SERGIO A.; GUTKIND, SILVIO; CURTI, CARLOS; LEOPOLDINO, ANDREIA M.. SET overexpression decreases cell detoxification efficiency: ALDH2 and GSTP1 are downregulated, DDR is impaired and DNA damage accumulates. FEBS Journal, v. 279, n. 24, p. 4615-4628, DEC 2012. (10/20536-4, 09/52228-0, 10/20384-0, 09/10783-7)