Role of hypoxia and autophagy in mda-mb-231 invasiveness

Manuela Indelicato, Bruna Pucci, Luana Schito, Valentina Reali, Michele Aventaggiato, Maria C. Mazzarino, Franca Stivala, Massimo Fini, Matteo A. Russo, Marco Tafani

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Survival strategies adopted by tumor cells in response to a hypoxic stress include activation of hypoxia-inducible factor 1 (HIF-1) and autophagy. However, the importance and the function of each molecular response is not well defined. In the present study, we investigated invasiveness, migration, matrix metalloproteinases (MMPs) activity, and cell survival of MDA-MB-231 cells under normoxia, hypoxia, and hypoxia/reoxygenation (H/R). Moreover, to assess the importance of hypoxia and autophagy on the parameters studied, cells were either left untreated or treated with Chetomin (a selective inhibitor of HIF-1α) or trifluoperazine (TFP, an activator of autophagy).Wefound that hypoxia and H/R stimulated invasiveness and migration of MDA-MB-231 cells with an increased MMP-2 activity. Chetomin and TFP differently regulated the cellular behavior under the oxygenation conditions studied. In fact, Chetomin was most effective in inhibiting cell invasion, MMPs activity, and cell survival under hypoxia but not normoxia or H/R. By contrast, TFP inhibition of cell invasion, migration, and cell survival was independent from oxygenation conditions. TFP-induced autophagy was inhibited by light chain protein 3 (LC3) silencing or 3-methyladenine (3MA) treatment. In fact, LC3-silenced cells were able to invade in the presence of TFP without any GATE16 processing and p62 degradation. Immunofluorescence assay showed that LC3 silencing inhibited TFP-induced autophagosome formation. However, we also showed that both TPF treatment and LC3 silencing caused cytoskeleton impairments suggesting a possible interaction between LC3 and cytoskeleton components. In conclusion, our study shows that hypoxia and autophagy by acting on common (HIF-1α) or separate (MMPs, cytoskeleton) targets differently regulate cell invasion, MMPs activity, and survival.

Original languageEnglish
Pages (from-to)359-368
Number of pages10
JournalJournal of Cellular Physiology
Volume223
Issue number2
DOIs
Publication statusPublished - May 2010

Fingerprint

Autophagy
Matrix Metalloproteinases
Hypoxia-Inducible Factor 1
Cells
Oxygenation
Light
Cytoskeleton
Proteins
Cell Survival
Trifluoperazine
Matrix Metalloproteinase 2
Cell Migration Inhibition
Tumors
Assays
Chemical activation
Hypoxia
Degradation
Fluorescent Antibody Technique
Processing
chetomin

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

Cite this

Indelicato, M., Pucci, B., Schito, L., Reali, V., Aventaggiato, M., Mazzarino, M. C., ... Tafani, M. (2010). Role of hypoxia and autophagy in mda-mb-231 invasiveness. Journal of Cellular Physiology, 223(2), 359-368. https://doi.org/10.1002/jcp.22041

Role of hypoxia and autophagy in mda-mb-231 invasiveness. / Indelicato, Manuela; Pucci, Bruna; Schito, Luana; Reali, Valentina; Aventaggiato, Michele; Mazzarino, Maria C.; Stivala, Franca; Fini, Massimo; Russo, Matteo A.; Tafani, Marco.

In: Journal of Cellular Physiology, Vol. 223, No. 2, 05.2010, p. 359-368.

Research output: Contribution to journalArticle

Indelicato, M, Pucci, B, Schito, L, Reali, V, Aventaggiato, M, Mazzarino, MC, Stivala, F, Fini, M, Russo, MA & Tafani, M 2010, 'Role of hypoxia and autophagy in mda-mb-231 invasiveness', Journal of Cellular Physiology, vol. 223, no. 2, pp. 359-368. https://doi.org/10.1002/jcp.22041
Indelicato M, Pucci B, Schito L, Reali V, Aventaggiato M, Mazzarino MC et al. Role of hypoxia and autophagy in mda-mb-231 invasiveness. Journal of Cellular Physiology. 2010 May;223(2):359-368. https://doi.org/10.1002/jcp.22041
Indelicato, Manuela ; Pucci, Bruna ; Schito, Luana ; Reali, Valentina ; Aventaggiato, Michele ; Mazzarino, Maria C. ; Stivala, Franca ; Fini, Massimo ; Russo, Matteo A. ; Tafani, Marco. / Role of hypoxia and autophagy in mda-mb-231 invasiveness. In: Journal of Cellular Physiology. 2010 ; Vol. 223, No. 2. pp. 359-368.
@article{5850989b841c4713903c6bf65f0fab56,
title = "Role of hypoxia and autophagy in mda-mb-231 invasiveness",
abstract = "Survival strategies adopted by tumor cells in response to a hypoxic stress include activation of hypoxia-inducible factor 1 (HIF-1) and autophagy. However, the importance and the function of each molecular response is not well defined. In the present study, we investigated invasiveness, migration, matrix metalloproteinases (MMPs) activity, and cell survival of MDA-MB-231 cells under normoxia, hypoxia, and hypoxia/reoxygenation (H/R). Moreover, to assess the importance of hypoxia and autophagy on the parameters studied, cells were either left untreated or treated with Chetomin (a selective inhibitor of HIF-1α) or trifluoperazine (TFP, an activator of autophagy).Wefound that hypoxia and H/R stimulated invasiveness and migration of MDA-MB-231 cells with an increased MMP-2 activity. Chetomin and TFP differently regulated the cellular behavior under the oxygenation conditions studied. In fact, Chetomin was most effective in inhibiting cell invasion, MMPs activity, and cell survival under hypoxia but not normoxia or H/R. By contrast, TFP inhibition of cell invasion, migration, and cell survival was independent from oxygenation conditions. TFP-induced autophagy was inhibited by light chain protein 3 (LC3) silencing or 3-methyladenine (3MA) treatment. In fact, LC3-silenced cells were able to invade in the presence of TFP without any GATE16 processing and p62 degradation. Immunofluorescence assay showed that LC3 silencing inhibited TFP-induced autophagosome formation. However, we also showed that both TPF treatment and LC3 silencing caused cytoskeleton impairments suggesting a possible interaction between LC3 and cytoskeleton components. In conclusion, our study shows that hypoxia and autophagy by acting on common (HIF-1α) or separate (MMPs, cytoskeleton) targets differently regulate cell invasion, MMPs activity, and survival.",
author = "Manuela Indelicato and Bruna Pucci and Luana Schito and Valentina Reali and Michele Aventaggiato and Mazzarino, {Maria C.} and Franca Stivala and Massimo Fini and Russo, {Matteo A.} and Marco Tafani",
year = "2010",
month = "5",
doi = "10.1002/jcp.22041",
language = "English",
volume = "223",
pages = "359--368",
journal = "Journal of cellular and comparative physiology",
issn = "0021-9541",
publisher = "Wiley-Liss Inc.",
number = "2",

}

TY - JOUR

T1 - Role of hypoxia and autophagy in mda-mb-231 invasiveness

AU - Indelicato, Manuela

AU - Pucci, Bruna

AU - Schito, Luana

AU - Reali, Valentina

AU - Aventaggiato, Michele

AU - Mazzarino, Maria C.

AU - Stivala, Franca

AU - Fini, Massimo

AU - Russo, Matteo A.

AU - Tafani, Marco

PY - 2010/5

Y1 - 2010/5

N2 - Survival strategies adopted by tumor cells in response to a hypoxic stress include activation of hypoxia-inducible factor 1 (HIF-1) and autophagy. However, the importance and the function of each molecular response is not well defined. In the present study, we investigated invasiveness, migration, matrix metalloproteinases (MMPs) activity, and cell survival of MDA-MB-231 cells under normoxia, hypoxia, and hypoxia/reoxygenation (H/R). Moreover, to assess the importance of hypoxia and autophagy on the parameters studied, cells were either left untreated or treated with Chetomin (a selective inhibitor of HIF-1α) or trifluoperazine (TFP, an activator of autophagy).Wefound that hypoxia and H/R stimulated invasiveness and migration of MDA-MB-231 cells with an increased MMP-2 activity. Chetomin and TFP differently regulated the cellular behavior under the oxygenation conditions studied. In fact, Chetomin was most effective in inhibiting cell invasion, MMPs activity, and cell survival under hypoxia but not normoxia or H/R. By contrast, TFP inhibition of cell invasion, migration, and cell survival was independent from oxygenation conditions. TFP-induced autophagy was inhibited by light chain protein 3 (LC3) silencing or 3-methyladenine (3MA) treatment. In fact, LC3-silenced cells were able to invade in the presence of TFP without any GATE16 processing and p62 degradation. Immunofluorescence assay showed that LC3 silencing inhibited TFP-induced autophagosome formation. However, we also showed that both TPF treatment and LC3 silencing caused cytoskeleton impairments suggesting a possible interaction between LC3 and cytoskeleton components. In conclusion, our study shows that hypoxia and autophagy by acting on common (HIF-1α) or separate (MMPs, cytoskeleton) targets differently regulate cell invasion, MMPs activity, and survival.

AB - Survival strategies adopted by tumor cells in response to a hypoxic stress include activation of hypoxia-inducible factor 1 (HIF-1) and autophagy. However, the importance and the function of each molecular response is not well defined. In the present study, we investigated invasiveness, migration, matrix metalloproteinases (MMPs) activity, and cell survival of MDA-MB-231 cells under normoxia, hypoxia, and hypoxia/reoxygenation (H/R). Moreover, to assess the importance of hypoxia and autophagy on the parameters studied, cells were either left untreated or treated with Chetomin (a selective inhibitor of HIF-1α) or trifluoperazine (TFP, an activator of autophagy).Wefound that hypoxia and H/R stimulated invasiveness and migration of MDA-MB-231 cells with an increased MMP-2 activity. Chetomin and TFP differently regulated the cellular behavior under the oxygenation conditions studied. In fact, Chetomin was most effective in inhibiting cell invasion, MMPs activity, and cell survival under hypoxia but not normoxia or H/R. By contrast, TFP inhibition of cell invasion, migration, and cell survival was independent from oxygenation conditions. TFP-induced autophagy was inhibited by light chain protein 3 (LC3) silencing or 3-methyladenine (3MA) treatment. In fact, LC3-silenced cells were able to invade in the presence of TFP without any GATE16 processing and p62 degradation. Immunofluorescence assay showed that LC3 silencing inhibited TFP-induced autophagosome formation. However, we also showed that both TPF treatment and LC3 silencing caused cytoskeleton impairments suggesting a possible interaction between LC3 and cytoskeleton components. In conclusion, our study shows that hypoxia and autophagy by acting on common (HIF-1α) or separate (MMPs, cytoskeleton) targets differently regulate cell invasion, MMPs activity, and survival.

UR - http://www.scopus.com/inward/record.url?scp=77950800322&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77950800322&partnerID=8YFLogxK

U2 - 10.1002/jcp.22041

DO - 10.1002/jcp.22041

M3 - Article

C2 - 20112292

AN - SCOPUS:77950800322

VL - 223

SP - 359

EP - 368

JO - Journal of cellular and comparative physiology

JF - Journal of cellular and comparative physiology

SN - 0021-9541

IS - 2

ER -