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Irradiation induces glioblastoma cell senescence and senescence-associated secretory phenotype

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Tumor Biology

Abstract

Glioblastoma multiforme (GBM) is one of the most aggressive and fatal primary brain tumors in humans. The standard therapy for the treatment of GBM is surgical resection, followed by radiotherapy and/or chemotherapy. However, the frequency of tumor recurrence in GBM patients is very high, and the survival rate remains poor. Delineating the mechanisms of GBM recurrence is essential for therapeutic advances. Here, we demonstrate that irradiation rendered 17–20 % of GBM cells dead, but resulted in 60–80 % of GBM cells growth-arrested with increases in senescence markers, such as senescence-associated beta-galactosidase-positive cells, H3K9me3-positive cells, and p53-p21CIP1-positive cells. Moreover, irradiation induced expression of senescence-associated secretory phenotype (SASP) mRNAs and NFκB transcriptional activity in GBM cells. Strikingly, compared to injection of non-irradiated GBM cells into immune-deficient mice, the co-injection of irradiated and non-irradiated GBM cells resulted in faster growth of tumors with the histological features of human GBM. Taken together, our findings suggest that the increases in senescent cells and SASP in GBM cells after irradiation is likely one of main reasons for tumor recurrence in post-radiotherapy GBM patients.

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Acknowledgments

We would like to thank all the members of the Cell Growth Regulation Laboratory for their helpful discussion and technical assistance. This work was supported by the National Nuclear Technology Program through the National Research Foundation (NRF) of Korea funded by the Ministry of Science, ICT, and Future Planning (No. 2013M2A2A7042530 to H. Kim), and a research grant (to S.Y. Oh) funded by the Institute of Life Science and Natural Resources at Korea University.

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Authors and Affiliations

  1. Department of Biotechnology, School of Life Sciences and Biotechnology, Korea University, Seoul, South Korea

    Hee-Young Jeon, Jun-Kyum Kim, Seok Won Ham & Hyunggee Kim

  2. Institute of Animal Molecular Biotechnology, Korea University, Seoul, South Korea

    Hee-Young Jeon, Jun-Kyum Kim & Hyunggee Kim

  3. Institute of Life Science and Natural Resources, Korea University, Seoul, South Korea

    Se-Yeong Oh

  4. Department of Biochemistry, Institute of Cell Differentiation and Aging, College of Medicine, Hallym University, Chuncheon, South Korea

    Jaebong Kim, Jae-Bong Park, Jae-Yong Lee & Sung-Chan Kim

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  1. Hee-Young Jeon
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Correspondence to Sung-Chan Kim or Hyunggee Kim.

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The study experiments were approved by the Animal Care Committee of the College of Life Sciences and Biotechnology, Korea University, according to government and institutional guidelines and regulations of Korea.

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sFig. 1

Irradiation-induced GBM cell death was decreased by inhibition of NFκB signaling. FACS analysis revealed that early apoptotic cell populations (Annexin V-positive/PI-negative) of IκBα mutant-expressing GBM cells (U87MG and LN229) decreased compared to control counterpart GBM cells on Day 3 after irradiation with 20 Gy. There was no obvious difference on necrotic cell population (Annexin V-negative/PI-positive) in these cells. (GIF 105 kb)

High Resolution Image (TIF 14313 kb)

sFig. 2

Irradiated GBM cells are not present in the tumor xenograft. a. FACS analysis showed cell populations expressing DsRed fluorescence in the mix of non-irradiated LN229 (1 × 10 ) and irradiated DsRed-expressing LN229 cells (2 × 10 ) before in vivo mouse co-injection. This experiment was set for a positive control. b. FACS analysis revealed that DsRed-positive cell populations were not present in tumors derived from the mix of non-irradiated LN229 and irradiated DsRed-expressing LN229 cells. Single cells dissociated from tumors derived from non-irradiated LN229 cells alone were used as a negative control. (GIF 133 kb)

High Resolution Image (TIF 17639 kb)

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Jeon, HY., Kim, JK., Ham, S.W. et al. Irradiation induces glioblastoma cell senescence and senescence-associated secretory phenotype. Tumor Biol. 37, 5857–5867 (2016). https://doi.org/10.1007/s13277-015-4439-2

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  • DOI: https://doi.org/10.1007/s13277-015-4439-2

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