Cellular and Molecular Neurobiology

, Volume 35, Issue 6, pp 785–795 | Cite as

The Internalization of Neurotensin by the Low-Affinity Neurotensin Receptors (NTSR2 and vNTSR2) Activates ERK 1/2 in Glioma Cells and Allows Neurotensin-Polyplex Transfection of tGAS1

  • Alberto E. Ayala-Sarmiento
  • Daniel Martinez-Fong
  • José Segovia
Original Research
First Online:
Received:
Accepted:

Abstract

Glioblastoma is the most malignant primary brain tumor and is very resistant to treatment; hence, it has a poor prognosis. Neurotensin receptor type 1 (NTSR1) plays a key role in cancer malignancy and has potential therapeutic applications. However, the presence and function of neurotensin (NTS) receptors in glioblastoma is not clearly established. RT-PCR assays showed that healthy (non-tumor) astroglial cells and C6 glioma cells express NTSR2 and its isoform (vNTSR2) rather than NTSR1. In glioma cells, NTS promotes the phosphorylation of extracellular signal-regulated kinases 1/2 (ERK 1/2), an effect that was completely abolished by blocking the internalization of the NTS/NTSR complex. We demonstrated pharmacologically that the internalization is dependent on the activation of NTSR2 receptors and it was prevented by levocabastine, a NTSR2 receptor antagonist. The internalization of NTSR2 and vNTSR2 was further demonstrated by its ability to mediate gene transfer (transfection) via the NTS-polyplex system. Expression of reporter transgenes and of the pro-apoptotic soluble form of growth arrest specific 1 (tGAS1) was observed in glioma cells. A significant reduction on the viability of C6 cells was determined when tGAS1 was transfected into glioma cells. Conversely, astroglial cells could neither internalize NTS nor activate ERK 1/2 and could not be transfected by the NTS-polyplex. These results demonstrate that the internalization process of NTSR2 receptors is a key regulator necessary to trigger the activation of the ERK 1/2. Our data support a new internalization pathway in glioma C6 cells that involve NTSR2/vNTSR2, which can be used to selectively transfer therapeutic genes using the NTS-polyplex system.

Keywords

Neurotensin receptor-2 Glioma Glia ERK 1/2 GAS1 Polyplex 
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Notes

Acknowledgments

This work was supported by Instituto de Ciencia y Tecnología del Gobierno del Distrito Federal (ICyTDF) Grant #ICYTDF/228/2010, ARN-CONACYT Grant #142947 (DMF), and Conacyt Grant #127357 (JS). AEAS was a recipient of CONACYT fellowship #244983.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Alberto E. Ayala-Sarmiento
    • 1
  • Daniel Martinez-Fong
    • 1
    • 2
  • José Segovia
    • 1
  1. 1.Departamento de Fisiología, Biofísica y NeurocienciasCentro de Investigación y de Estudios Avanzados del IPNMexicoMexico
  2. 2.Programa de Nanociencias y NanotecnologíaCentro de Investigación y de Estudios Avanzados del Instituto Politécnico NacionalMexicoMexico

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