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Combinatorial therapy with adenoviral-mediated PTEN and a PI3K inhibitor suppresses malignant glioma cell growth in vitro and in vivo by regulating the PI3K/AKT signaling pathway

  • Original Article – Cancer Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

Glioblastoma is a highly invasive and challenging tumor of the central nervous system. The mutation/deletion of the tumor suppressor phosphatase and tensin homolog (PTEN) gene is the main genetic change identified in glioblastomas. PTEN plays a critical role in tumorigenesis and has been shown to be an important therapeutic target. The phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 is commonly used to inhibit glioma cell growth via regulation of the PI3K/AKT signaling pathway. In this study, we examined the growth inhibitory effects of a combinatorial therapy of adenoviral-mediated PTEN (Ad-PTEN) and LY294002 on LN229 and U251 glioma cells in vitro and on tumor xenografts in vivo.

Methods

In vitro, LN229 and U251 glioma cells were treated by combinatorial therapy with Ad-PTEN and LY294002. The growth ability was determined by MTT assay. The cell cycle distribution was analyzed by flow cytometry. Cell invasive ability was analyzed by transwell invasion assay and cell apoptosis analysis via FITC-Annexin V analysis. In vivo, U251 subcutaneous glioblastoma xenograft was used to assay anti-tumor effect of combinatorial therapy with Ad-PTEN and LY294002 by mean volume of tumors, immunohistochemistry and TUNEL method.

Results

The combinatorial treatment clearly suppressed cell proliferation, arrested the cell cycle, reduced cell invasion and promoted cell apoptosis compared with the Ad-PTEN or LY294002 treatment alone. The treatment worked by inhibiting the PI3K/AKT pathway. In addition, the growth of U251 glioma xenografts treated with the combination of Ad-PTEN and LY294002 was significantly inhibited compared with those treated with Ad-PTEN or LY294002 alone.

Conclusions

Our data indicated that the combination of Ad-PTEN and LY294002 effectively suppressed the malignant growth of human glioma cells in vitro and in tumor xenografts, suggesting a promising new approach for glioma gene therapy that warrants further investigation.

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Abbreviations

PTEN:

Phosphatase and tensin homolog

PI3K:

Phosphatidylinositol 3-kinase

AKT:

AKT protein family, whose members are also called protein kinases B

DMEM:

Dulbecco’s Modified Eagle’s medium

DMSO:

Dimethylsulfoxide

PBS:

Phosphate-buffered saline

References

  • Becker KP, Yu J (2012) Status quo-standard-of-care medical and radiation therapy for glioblastoma. Cancer J 18:12–19. doi:10.1097/PPO.0b013e318244d7eb

    Article  CAS  PubMed  Google Scholar 

  • Bermúdez Brito M, Goulielmaki E, Papakonstanti EA (2015) Focus on PTEN regulation. Front Oncol 5:166. doi:10.3389/fonc.2015.00166

    Article  PubMed  PubMed Central  Google Scholar 

  • Chautard E, Loubeau G, Tchirkov A, Chassagne J, Vermot-Desroches C, Morel L, Verrelle P (2010) Akt signaling pathway: a target for radiosensitizing human malignant glioma. Neuro Oncol 12:434–443. doi:10.1093/neuonc/nop059

    CAS  PubMed  PubMed Central  Google Scholar 

  • Dressing GE, Knutson TP, Schiewer MJ, Daniel AR, Hagan CR, Diep CH, Knudsen KE, Lange CA (2014) Progesterone receptor-cyclin D1 complexes induce cell cycle-dependent transcriptional programs in breast cancer cells. Mol Endocrinol 28:442–457. doi:10.1210/me.2013-1196

    Article  PubMed  PubMed Central  Google Scholar 

  • Fu XY, Zhang S, Wang K, Yang MF, Fan CD, Sun BL (2015) Caudatin inhibits human glioma cells growth through triggering DNA damage-mediated cell cycle arrest. Cell Mol Neurobiol 35:953–959. doi:10.1007/s10571-015-0190-x

    Article  CAS  PubMed  Google Scholar 

  • Gabrusiewicz K, Liu D, Cortes-Santiago N, Hossain MB, Conrad CA, Aldape KD, Fuller GN, Marini FC, Alonso MM, Idoate MA, Gilbert MR, Fueyo J, Gomez-Manzano C (2014) Anti-vascular endothelial growth factor therapy-induced glioma invasion is associated with accumulation of Tie2-expressing monocytes. Oncotarget 5:2208–2220. doi:10.18632/oncotarget.1893

    Article  PubMed  PubMed Central  Google Scholar 

  • Han L, Zhang AL, Xu P, Yue X, Yang Y, Wang GX, Jia ZF, Pu PY, Kang CS (2010) Combination gene therapy with PTEN and EGFR siRNA suppresses U251 malignant glioma cell growth in vitro and in vivo. Med Oncol 27:843–852. doi:10.1007/s12032-009-9295-8

    Article  CAS  PubMed  Google Scholar 

  • Jiang Z, Pore N, Cerniglia GJ, Mick R, Georgescu MM, Bernhard EJ, Hahn SM, Gupta AK, Maity A (2007) Phosphatase and tensin homologue deficiency in glioblastoma confers resistance to radiation and temozolomide that is reversed by the protease inhibitor nelfinavir. Cancer Res 67:4467–4473. doi:10.1158/0008-5472.CAN-06-3398

    Article  CAS  PubMed  Google Scholar 

  • Jiang L, Wang C, Lei F, Zhang LJ, Zhang X, Liu AB, Wu GY, Zhu JR, Song L (2015) miR-93 promotes cell proliferation in gliomas through activation of PI3K/Akt signaling pathway. Oncotarget 6:8286–8299. doi:10.18632/oncotarget.3221

    Article  PubMed  PubMed Central  Google Scholar 

  • Lee CM, Fuhrman CB, Planelles V, Peltier MR, Gaffney DK, Soisson AP, Dodson MK, Tolley HD, Green CL, Zempolich KA (2006) Phosphatidylinositol 3-kinase inhibition by LY294002 radiosensitizes human cervical cancer cell lines. Clin Cancer Res 12:250–256. doi:10.1158/1078-0432.CCR-05-1084

    Article  CAS  PubMed  Google Scholar 

  • Malmström A, Grønberg BH, Marosi C, Stupp R, Frappaz D, Schultz H, Abacioglu U, Tavelin B, Lhermitte B, Hegi ME, Rosell J, Henriksson R, Nordic Clinical Brain Tumour Study Group (NCBTSG), for the Nordic Clinical Brain Tumour Study Group (NCBTSG) (2012) Temozolomide versus standard 6-week radiotherapy versus hypofractionated radiotherapy in patients older than 60 years with glioblastoma: the Nordic randomised, phase 3 trial. Lancet Oncol 13:916–926. doi:10.1016/S1470-2045(12)70265-6

    Article  PubMed  Google Scholar 

  • Miao Y, Zheng W, Li N, Su Z, Zhao L, Zhou H, Jia L (2017) MicroRNA-130b targets PTEN to mediate drug resistance and proliferation of breast cancer cells via the PI3K/Akt signaling pathway. Sci Rep 7:41942. doi:10.1038/srep41942

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nakamura JL, Karlsson A, Arvold ND, Gottschalk AR, Pieper RO, Stokoe D, Haas-Kogan DA (2005) PKB/Akt mediates radiosensitization by the signaling inhibitor LY294002 in human malignant gliomas. J Neurooncol 71:215–222. doi:10.1007/s11060-004-1718-y

    Article  CAS  PubMed  Google Scholar 

  • Okura H, Smith CA, Rutka JT (2014) Gene therapy for malignant glioma. Mol Cell Ther 2:21

    Article  PubMed  PubMed Central  Google Scholar 

  • Opel D, Westhoff MA, Bender A, Braun VD, Debatin KM, Fulda S (2008) Phosphatidylinositol 3-kinase inhibition broadly sensitizes glioblastoma cells to death receptor- and drug-induced apoptosis. Cancer Res 68:6271–6280. doi:10.1158/0008-5472.CAN-07-6769

    Article  CAS  PubMed  Google Scholar 

  • Pu PY, Kang CS, Li J, Jiang H, Cheng JQ (2006) The effects of antisense AKT2 RNA on the inhibition of malignant glioma cell growth in vitro and in vivo. J Neurooncol 76:1–11. doi:10.1007/s11060-005-3029-3

    Article  CAS  PubMed  Google Scholar 

  • Ren Y, Zhou X, Mei M, Yuan XB, Han L, Wang GX, Jia ZF, Xu P, Pu PY, Kang CS (2010) MicroRNA-21 inhibitor sensitizes human glioblastoma cells U251 (PTEN-mutant) and LN229 (PTEN-wild type) to Taxol. BMC Cancer 10:27. doi:10.1186/1471-2407-10-27

    Article  PubMed  PubMed Central  Google Scholar 

  • Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, Ludwin SK, Allgeier A, Fisher B, Belanger K, Hau P, Brandes AA, Gijtenbeek J, Marosi C, Vecht CJ, Mokhtari K, Wesseling P, Villa S, Eisenhauer E, Gorlia T, Weller M, Lacombe D, Cairncross JG, Mirimanoff RO, European Organisation for Research and Treatment of Cancer Brain Tumour and Radiation Oncology Groups (2009) Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol 10:459–466. doi:10.1016/S1470-2045(09)70025-7

    Article  CAS  PubMed  Google Scholar 

  • Tan Y, Huang N, Zhang X, Hu J, Cheng S, Pi L, Cheng Y (2016) KIAA0247 suppresses the proliferation, angiogenesis and promote apoptosis of human glioma through inactivation of the AKT and Stat3 signaling pathway. Oncotarget 7:87100–87113. doi:10.18632/oncotarget.13527

    PubMed  PubMed Central  Google Scholar 

  • Thomas S, Quinn BA, Das SK, Dash R, Emdad L, Dasgupta S, Wang XY, Dent P, Reed JC, Pellecchia M, Sarkar D, Fisher PB (2013) Targeting the Bcl-2 family for cancer therapy. Expert Opin Ther Targets 17:61–75. doi:10.1517/14728222.2013.733001

    Article  CAS  PubMed  Google Scholar 

  • Wang XR, Luo H, Li HL, Cao L, Wang XF, Yan W, Wang YY, Zhang JX, Jiang T, Kang CS, Liu N, You YP, Chinese Glioma Cooperative Group (CGCG) (2013) Overexpressed let-7a inhibits glioma cell malignancy by directly targeting K-ras, independently of PTEN. Neuro Oncol 15:1491–1501. doi:10.1093/neuonc/not107

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wen PY, Reardon DA (2016) Neuro-oncology in 2015: progress in glioma diagnosis, classification and treatment. Nat Rev Neurol 12:69–70. doi:10.1038/nrneurol.2015.242

    Article  CAS  PubMed  Google Scholar 

  • Wu YL, Maachani UB, Schweitzer M, Singh R, Wang M, Chang R, Souweidane MM (2017) Dual inhibition of PI3K/AKT and MEK/ERK pathways induces synergistic antitumor effects in diffuse intrinsic pontine glioma cells. Transl Oncol 10:221–228. doi:10.1016/j.tranon.2016.12.008

    Article  PubMed  PubMed Central  Google Scholar 

  • Xu P, Zhang AL, Jiang RC, Qiu MZ, Kang CS, Jia ZF, Wang GX, Han L, Fan X, Pu PY (2013) The different role of Notch1 and Notch2 in astrocytic gliomas. PLoS One 8:e53654. doi:10.1371/journal.pone.0053654

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yao C, Li P, Song H, Song F, Qu Y, Ma X, Shi R, Wu J (2016) CXCL12/CXCR4 axis upregulates twist to induce EMT in human glioblastoma. Mol Neurobiol 53:3948–3953. doi:10.1007/s12035-015-9340-x

    Article  CAS  PubMed  Google Scholar 

  • Yazdani Y, Sharifi Rad MR, Taghipour M, Chenari N, Ghaderi A, Razmkhah M (2016) Genistein suppression of matrix metalloproteinase 2 (MMP-2) and vascular endothelial growth factor (VEGF) expression in mesenchymal stem cell like cells isolated from high and low grade gliomas. Asian Pac J Cancer Prev 17:6203–6207. doi:10.22034/APJCP.2016.17.6203

    Google Scholar 

  • Zhang CZ, Zhang JX, Hao JW, Shi ZD, Wang YY, Han L, Yu SZ, You YP, Jiang T, Wang JH, Liu ML, Pu PY, Kang CS (2012) High level of miR-221/222 confers increased cell invasion and poor prognosis in glioma. J Transl Med 10:119. doi:10.1186/1479-5876-10-119

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang FY, Hu Y, Que ZY, Wang P, Liu YH, Wang ZH, Xue YX (2015) Shikonin inhibits the migration and invasion of human glioblastoma cells by targeting phosphorylated β-catenin and phosphorylated PI3K/Akt: a potential mechanism for the anti-glioma efficacy of a traditional Chinese herbal medicine. Int J Mol Sci 16:23823–23848. doi:10.3390/ijms161023823

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou X, Ren Y, Moore L, Mei M, You YP, Xu P, Wang BL, Wang GX, Jia ZF, Pu PY, Zhang W, Kang CS (2010) Downregulation of miR-21 inhibits EGFR pathway and suppresses the growth of human glioblastoma cells independent of PTEN status. Lab Invest 90:144–155. doi:10.1038/labinvest.2009.126

    Article  CAS  PubMed  Google Scholar 

  • Zhu D, Tu M, Zeng B, Cai L, Zheng W, Su Z, Yu Z (2017) Up-regulation of miR-497 confers resistance to temozolomide in human glioma cells by targeting mTOR/Bcl-2. Cancer Med 6:452–462. doi:10.1002/cam4.987

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by the China National Natural Scientific Fund (81572490) and the Tianjin Science and Technology Committee (13JCZDJC31000).

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Authors and Affiliations

  1. Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin, 300052, People’s Republic of China

    Yang Nan, Liyun Guo, Yunpeng Song, Le Wang, Kai Yu, Qiang Huang & Yue Zhong

  2. Laboratory of Neuro-Oncology, Tianjin Neurological Institute, Tianjin, 300052, People’s Republic of China

    Yang Nan, Yunpeng Song, Le Wang & Yue Zhong

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  1. Yang Nan
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  2. Liyun Guo
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  3. Yunpeng Song
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  4. Le Wang
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  5. Kai Yu
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  6. Qiang Huang
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  7. Yue Zhong
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Correspondence to Yue Zhong.

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All authors declare that they have no conflict of interest. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

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Nan, Y., Guo, L., Song, Y. et al. Combinatorial therapy with adenoviral-mediated PTEN and a PI3K inhibitor suppresses malignant glioma cell growth in vitro and in vivo by regulating the PI3K/AKT signaling pathway. J Cancer Res Clin Oncol 143, 1477–1487 (2017). https://doi.org/10.1007/s00432-017-2415-5

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