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Γ-Ionizing radiation-induced activation of the EGFR–p38/ERK–STAT3/CREB-1–EMT pathway promotes the migration/invasion of non-small cell lung cancer cells and is inhibited by podophyllotoxin acetate

Tumor Biology volume 37, pages 7315–7325 (2016)Cite this article

Abstract

Here, we report a new intracellular signaling pathway involved in γ-ionizing radiation (IR)-induced migration/invasion and show that podophyllotoxin acetate (PA) inhibits the IR-induced invasion and migration of A549 cells (a non-small cell lung cancer (NSCLC) cell line). Our results revealed that IR increased the invasion/migration of A549 cells, and this effect was decreased by 10 nM PA treatment. PA also inhibited the expressions/activities of matrix metalloprotase (MMP) -2, MMP-9, and vimentin, suggesting that PA could block the IR-induced epithelial-mesenchymal transition (EMT). The IR-induced increases in invasion/migration were associated with the activation of EGFR-AKT, and PA inhibited this effect. P38 and p44/42 ERK were also involved in IR-induced invasion/migration, and combined treatments with PA plus inhibitors of each MAPK synergistically blocked this invasion/migration. In terms of transcription factors (TFs), IR-induced increases in cyclic AMP response element-binding protein-1 (CREB-1) and signal transducer and activator of transcription 3 (STAT3) increased invasion/migration and EMT. PA also inhibited these transcription factors and then blocked IR-induced invasion/migration. Collectively, these results indicate that IR induces cancer cell invasion/migration by activating the EGFR–p38/ERK–CREB-1/STAT3–EMT pathway and that PA blocks this pathway to inhibit IR-induced invasion/migration.

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Acknowledgments

This work was supported by a grant from the Nuclear Research & Development Program of the National Research Foundation of Korea (NRF), which is funded by the Korean government (MEST; 2012M2A2A7010422), and by the Basic Science Research Program of the NRF (NRF-2014R1A1A2054985).

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Affiliations

  1. Department of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, 215-4, Gongneung-Dong, Nowon-Gu, Seoul, 139-706, South Korea

    Jeong Hyun Cho, Wan Gi Hong, Sang-Gu Hwang, Hong-Duck Um & Jong Kuk Park

  2. Department of Biological Sciences, Gangwon National University, Hyoja 2-dong, Chuncheon-si, Gangwon-do, 200-701, South Korea

    Jeong Hyun Cho & Yu-Jin Jung

  3. Department of Biological Science, College of Biological Science, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 05029, South Korea

    Jaeseok Lee

  4. Musculoskeletal Bio-Organ Center, College of Life Science, Kyung Hee University, 1732 Deogyeongdaero Giheung-gu, Yongin-si, Gyeonggi-do, 446-701, South Korea

    Eunah Lee

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  1. Jeong Hyun Cho
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  2. Wan Gi Hong
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Correspondence to Jong Kuk Park.

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Supplementary Fig. 1

Gelatin zymography assay to detect activity of MMPs. Cells were treated with various combinations of IR, PA, SB (10 μM SB203580), PD (10 μM PD98059) and Gefinitib (10 μM), and gelatin zymography was used to detect the activities of MMP-2 a and -9 b. Representative data from triplicate experiments are shown. (PPTX 116 kb)

Supplementary Fig. 2

Apoptotic cell death detection with PI uptake. IR increases invasion/migration by activating STAT3, and these effects are modulated by PA. Cells were treated with IR and C188-9 (10 μM), and apoptotic cell death was determined by PI-uptake assay. Representative data from triplicate experiments are shown. (PPTX 146 kb)

Supplementary Fig. 3

Apoptotic cell death detection with PI uptake. IR increases invasion/migration by activating CREB-1, and these effects are modulated by PA. Cells were subjected to IR and treated with an inhibitor of the CBP-CREB interaction (CREB inh; 10 μM), and apoptotic cell death was determined by PI-uptake assay. Representative data from repeated experiments performed in triplicate are shown. (PPTX 153 kb)

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Cho, J.H., Hong, W.G., Jung, YJ. et al. Γ-Ionizing radiation-induced activation of the EGFR–p38/ERK–STAT3/CREB-1–EMT pathway promotes the migration/invasion of non-small cell lung cancer cells and is inhibited by podophyllotoxin acetate. Tumor Biol. 37, 7315–7325 (2016). https://doi.org/10.1007/s13277-015-4548-y

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Keywords

  • Radiation
  • Podophyllotoxin acetate
  • Invasion
  • Migration
  • NSCLC