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Journal of Neuro-Oncology

, Volume 130, Issue 1, pp 43–52 | Cite as

The role of AKT isoforms in glioblastoma: AKT3 delays tumor progression

  • Anna JoyEmail author
  • Manisha Kapoor
  • Joseph Georges
  • Lacy Butler
  • Yongchang Chang
  • Chaokun Li
  • Acacia Crouch
  • Ivan Smirnov
  • Mitsitoshi Nakada
  • James Hepler
  • Max Marty
  • Burt G. Feuerstein
Laboratory Investigation

Abstract

The growth factor receptor/PI3K/AKT pathway is an important drug target in many cancers including Glioblastoma. AKT, a key node in the pathway, has 3 isoforms, AKT1, AKT2 and AKT3. Here we investigate their role in GBM. We find each activated, ser473 phosphorylated isoform is present in some GBMs but expression patterns vary. There is a direct relationship between human GBM patient outcome and both AKT1 and AKT2 mRNA levels, but an inverse relationship with AKT3 mRNA. Furthermore, AKT3 mRNA levels were high in a less aggressive GBM subtype. Overexpressing AKT3 improves survival in a rodent model of GBM and decreases colony forming efficiency, but not growth rate, in glioma cells. Silencing AKT3 slows cell cycle progression in one cell line and increases apoptosis in another. Our studies of AKT3 substrates indicate (1) silencing both AKT2 and AKT3 reduces GSK3 phosphorylation (2) only AKT2 silencing reduces S6 phosphorylation. Since S6 phosphorylation is a marker of mTORC1 activity this indicates that AKT2 activates mTORC1, but AKT3 does not. Our results indicate AKT isoforms have different roles and downstream substrates in GBM. Unexpectedly, they indicate AKT3 delays tumor progression. Therefore strategies that inhibit AKT3 may be unhelpful in some GBM patients.

Keywords

Glioblastoma GBM AKT AKT1 AKT2 AKT3 

Notes

Acknowledgments

This work was supported by KO1 NS064952 from the National Institutes of Health to AMJ, by 1R21EB020237 from the National Institutes of Health and an award from the Bruce Halle foundation to BGF and Barrow Neurological Foundation awards to AMJ and BGF.

Funding

This study was funded by the National Institute of Neurological Disorders and Stroke at the National Institutes of Health (K01 NS064952 to A.J.); by the National Institute of Biomedical Imaging and Bioengineering at the National Institutes of Health (R21 EB020237 to BGF); and by the Barrow Neurological Foundation and Diane and Bruce Halle Fund (B.G.F. and A.J.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

Ethical approval human All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional review board and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Ethical approval animal All applicable international, national, and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were approved by our Institutional Animal Care and Use Committee (IACUC) and were in accordance with the ethical standards of the institution at which the studies were conducted.

Informed consent

For this type of study formal consent is not required.

Supplementary material

11060_2016_2220_MOESM1_ESM.pdf (183 kb)
Supplementary material 1 (PDF 182 KB)
11060_2016_2220_MOESM2_ESM.tif (718 kb)
Supplemental Fig S1 AKT3V1 is the predominant alternately spliced mRNA in GBM cell lines. (a) Schematic illustration of the domain structure of AKT3V1 and AKT3V2. (b) QRT-PCR analysis of AKT3V1 and AKT3V2 mRNA in GBM cell lines. (TIF 718 KB)
11060_2016_2220_MOESM3_ESM.tif (381 kb)
Supplemental Fig S2 Effect of silencing each AKT isoform on flow cytometric analysis of DNA content in PI stained glioma cells. (a) U87 and (b) U251 cells were incubated for 3 days after exposure to the specified AKT isoform siRNA then the histogram of count number vs PI fluorescence obtained. (a) % cells in each phase of the cell cycle was determined using FCS express. Percent cells with DNA content greater than G2/M in U87 cells is indicated. (b) Percent U251 cells in subG0 peak is indicated. (TIF 380 KB)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Anna Joy
    • 1
    • 5
    Email author
  • Manisha Kapoor
    • 1
  • Joseph Georges
    • 1
  • Lacy Butler
    • 1
  • Yongchang Chang
    • 1
  • Chaokun Li
    • 1
  • Acacia Crouch
    • 1
  • Ivan Smirnov
    • 2
  • Mitsitoshi Nakada
    • 3
  • James Hepler
    • 1
  • Max Marty
    • 1
  • Burt G. Feuerstein
    • 4
  1. 1.St. Josephs Hospital and Medical CenterPhoenixUSA
  2. 2.University of California at San FranciscoSan FranciscoUSA
  3. 3.Department of NeurosurgeryKanazawa UniversityKanazawaJapan
  4. 4.Department of NeurologyUniversity of Arizona College of MedicinePhoenixUSA
  5. 5.PhoenixUSA

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