Skip to main content

Advertisement

SpringerLink
Log in

Endostatin exerts radiosensitizing effect in non-small cell lung cancer cells by inhibiting VEGFR2 expression

  • Research Article
  • Published:
Clinical and Translational Oncology Aims and scope Submit manuscript

Abstract

Background

To determine the effects of endostatin on vascular growth factor receptor 2 (VEGFR2) expression in non-small cell lung cancer (NSCLC) cells and the mechanisms underlying its radiosensitizing effect.

Methods

VEGFR2 mRNA levels were determined in different NSCLC cell lines using qRT-PCR. RT-PCR and Western blot assays were used to assess the expression of mRNA and proteins. The radiosensitivity of the cells was determined by colony-formation assays; and cell apoptosis and cell cycle distribution were determined by flow cytometry.

Results

VEGFR2 mRNA levels differed among the five NSCLC cell lines (P < 0.01), with the highest expression in Calu-1 cells and lowest in A549 cells. Endostatin significantly inhibited the growth of Calu-1 cells (P < 0.01) (IC20 = 296.5 μg/ml), and the expression of VEGFR2 and HIF-1α (P < 0.05). Phosphorylation of protein kinase B (Akt), extracellular signal-regulated kinases 1/2 (ERK1/2), and p38 were significantly lower in endostatin-treated cells than control (P < 0.05). Endostatin enhanced the radiosensitivity of Calu-1 cells to SER = 1.38 and induced apoptosis (P < 0.01) and G2/M blockage (P < 0.01). However, endostatin had limited effects on A549 cells. Compared with Calu-1 cells, there was not significantly effects on cell radiosensitivity (SER = 1.09).

Conclusions

Endostatin induces apoptosis and enhances radiosensitivity of the VEGFR2 high-expressing cell line Calu-1, but it has a limited effect on the VEGFR2 low-expressing cell line A549.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Yeo SG, Kim ES. Efficient approach for determining four-dimensional computed tomography-based internal target volume in stereotactic radiotherapy of lung cancer[J]. Radiat Oncol J. 2013;31(4):247–51.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Chi A, Liao Z, Nguyen NP, Xu J, Stea B, Komaki R. Systemic review of the patterns of failure following stereotactic body radiation therapy in early-stage non-small-cell lung cancer: clinical implications. Radiother Oncol. 2010;94:1–11.

    Article  PubMed  Google Scholar 

  3. Langendorff H, Koch R. Studies on biological radioprotection. IX. Effect of SH-blockers on radiosensitivity. Strahlentherapie. 1954;95(4):535–41.

    PubMed  CAS  Google Scholar 

  4. Jiang X-D, Qiao Y, Dai P, Wu J, Song D-A, Li S-Q, et al. Preliminary clinical study of weekly recombinant human endostatin as a hypoxic tumour cell radiosensitiser combined with radiotherapy in the treatment of NSCLC. Clin Transl Oncol. 2012;14(6):465–70.

    Article  PubMed  CAS  Google Scholar 

  5. Jiang X-D, Ding M-H, Qiao Y, Liu Y, Liu L. Study on lung cancer cells expressing VEGFR2 and the impact on the effect of RHES combined with radiotherapy in the treatment of brain metastases. Clin Lung Cancer. 2014;15(2):e23–9.

    Article  PubMed  Google Scholar 

  6. Donnem T, Al-Shibli K, Andersen S, Al-Saad S, Busund L-T, Bremnes RM. Combination of low vascular endothelial growth factor A (VEGF-A)/VEGF receptor 2 expression and high lymphocyte infiltration is a strong and independent favorable prognostic factor in patients with nonsmall cell lung cancer. Cancer. 2010;116:4318–25.

    Article  PubMed  CAS  Google Scholar 

  7. Dhakal HP, Naume B, Synnestvedt M, Borgen E, Kaaresen R, Schlichting E, et al. Expression of vascular endothelial growth factor and vascular endothelial growth factor receptors 1 and 2 in invasive breast carcinoma: prognostic significance and relationship with markers for aggressiveness. Histopathology. 2012;61:350–64.

    Article  PubMed  Google Scholar 

  8. Grosso S, Doyen J, Parks SK, Bertero T, Paye A, Cardinaud B, et al. MiR-210 promotes a hypoxic phenotype and increases radioresistance in human lung cancer cell lines. Cell Death Dis. 2013;4:e544.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  9. Yang F, Tang X, Riquelme E, Behrens B, Nilsson MB, Giri U, Tang X, et al. Increased VEGFR-2 gene copy is associated with chemoresistance and shorter survival in patients with non-small-cell lung carcinoma who receive adjuvant chemotherapy. Cancer Res. 2011;71(16):5512–21.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  10. Semenza G. Signal transduction to hypoxia-inducible factor 1. Biochem Pharmacol. 2002;64(5–6):993–8.

    Article  PubMed  CAS  Google Scholar 

  11. Kerbel RS. Tumor angiogenesis. N Engl J Med. 2008;358(19):2039–49.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  12. Kim Y-M, Hwang S, Kim Y-M, Pyun B-J, Kim T-Y, Lee S-T, et al. Endostatin blocks vascular endothelial growth factor-mediated signaling via direct interaction with KDR/Flk-1.[J]. J Biol Chem. 2002;277(31):27872–9.

    Article  PubMed  CAS  Google Scholar 

  13. Brown JM, Giaccia AJ. The unique physiology of solid tumors: opportunities(and problems)for cancer therapy[J]. Cancer Res. 1998;58(7):1408–16.

    PubMed  CAS  Google Scholar 

  14. Meijer TWH, Kaanders JHAM, Span PN, Bussink J. Targeting hypoxia, HIF-1, and tumor glucose metabolism to improve radiotherapy efficacy. Clin Cancer Res. 2012;18(20):5585–94.

    Article  PubMed  CAS  Google Scholar 

  15. Agani F, Jiang B-H. Oxygen-independent regulation of HIF-1: novel involvement of PI3 K/AKT/mTOR pathway in cancer. Curr Cancer Drug Targets. 2013;13:245–51.

    Article  PubMed  CAS  Google Scholar 

  16. Karapetsas A, Giannakakis A, Pavlaki M, Panayiotidis M, Sandaltzopoulos R, Alex Galanis. Biochemical and molecular analysis of the interaction between ERK2 MAP kinase and hypoxia inducible factor-1α. Int J Biochem Cell Biol. 2011;43(11):1582–90.

    Article  PubMed  CAS  Google Scholar 

  17. Sodhi A, Montaner S, Patel V, Zohar M, Bais C, Mesri EA, et al. The Kaposi’s sarcoma-associated herpes virus G protein-coupled receptor up-regulates vascular endothelial growth factor expression and secretion through mitogen-activated protein kinase and p38 pathways acting on hypoxia-inducible factor 1alpha. Cancer Res. 2000;60(17):4873–80.

    PubMed  CAS  Google Scholar 

  18. Sang N, Stiehl DP, Bohensky J, Leshchinsky I, Srinivas V, Caro J. MAPK signaling up-regulates the activity of hypoxia-inducible factors by its effects on p300. J Biol Chem. 2003;278(16):14013–9.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  19. Nilsson MB, Zage PE, Zeng L, Xu L, Cascone T, Wu HK, et al. Multiple receptor tyrosine kinases regulate HIF-1alpha and HIF-2alpha in normoxia and hypoxia in neuroblastoma: implications for antiangiogenic mechanisms of multikinase inhibitors. Oncogene. 2010;29(20):2938–49.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by the National Natural Science Foundation of China (NO. 81472792), Research Fund from Ministry of Health (W201210), and the National Natural Science Foundation of Jiangsu Province (BK2012661).

Conflict of interests

We have read and understood Clinical and Translational Oncology’s policy on disclosing conflicts of interest and declare that we have none.

Ethics statement

Our study did not refer to clinical trial and animal trial, so there was no ethics statement.

Author information

Author notes

    Authors and Affiliations

    1. Xuzhou Medical College Graduate Academy, Xuzhou, 221006, China

      L. Liu, Y. Liu, B. Liu & H. Chen

    2. Department of Radiation Oncology, Lianyungang First People’s Hospital, No. 182, Tongguan Road, Lianyungang, 222002, Jiangsu, China

      Y. Qiao, C. Hu, Y. Xia, L. Wang & X. Jiang

    Authors
    1. L. Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Y. Qiao
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. C. Hu
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Y. Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Y. Xia
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. L. Wang
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. B. Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. H. Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. X. Jiang
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to X. Jiang.

    Additional information

    L. Liu, Y. Qiao and C. Hu contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Liu, L., Qiao, Y., Hu, C. et al. Endostatin exerts radiosensitizing effect in non-small cell lung cancer cells by inhibiting VEGFR2 expression. Clin Transl Oncol 18, 18–26 (2016). https://doi.org/10.1007/s12094-015-1319-6

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s12094-015-1319-6

    Keywords

    Search

    Navigation