Surgical Endoscopy

, Volume 32, Issue 3, pp 1533–1539 | Cite as

Simvastatin enhances radiation sensitivity of colorectal cancer cells

  • Georgios Karagkounis
  • Jennifer DeVecchio
  • Sylvain Ferrandon
  • Matthew F. Kalady
Article
  • 152 Downloads

Abstract

Introduction

Neoadjuvant chemoradiation (CRT) for rectal cancer induces variable responses, and better response has been associated with improved oncologic outcomes. Our group has previously shown that the administration of HMG-CoA reductase inhibitors, commonly known as statins, is associated with improved response to neoadjuvant CRT in rectal cancer patients. The purpose of this study was to study the effects of simvastatin on colorectal cancer cells and explore its potential as a radiation-sensitizer in vitro.

Methods

Four colorectal cancer cell lines (SW480, DLD1, SW837, and HRT18) were used to test the effects of simvastatin alone, radiation alone, and combination therapy. Outcome measures included ATP-based cell viability, colony formation, and protein (immunoblot) assays.

Results

The combination of radiation and simvastatin inhibited colony formation and cell viability of all four CRC lines, to a greater degree than either treatment alone (p < 0.01). In addition, the effects of simvastatin in this combination therapy were dose dependent, with increased concentrations resulting in more potentiated inhibitory effects. The radiosensitizing effects of simvastatin on cell viability were negated by the presence of exogenous GGPP in the media. On protein analyses of irradiated cells, simvastatin treatment inhibited phosphorylation of ERK1/2, in a dose-dependent manner, while the total levels of ERK1/2 remained stable. In addition, the combined treatment resulted in increased levels of cleaved caspase 3, indicating greater apoptotic activity in the cells treated with radiation and simvastatin together.

Conclusions

Treatment with simvastatin hindered CRC cell viability and enhanced radiation sensitivity in vitro. These effects were tied to the depletion of GGPP and the decreased phosphorylation of ERK1/2, suggesting a prominent role for the EGFR-RAS-ERK1/2 pathway, through which statin enhances radiation sensitivity.

Keywords

Colorectal cancer Radiation Sensitivity Response Statins 

Notes

Acknowledgements

Georgios Karagkounis is the recipient of the 2015 Society of American Gastrointestinal and Endoscopic Surgeons (SAGES) Career Development Award (CDA) and the American Society of Colon and Rectal Surgeons (ASCRS) General Surgery Resident Research Initiation Grant #25. Matthew F. Kalady is the Krause-Lieberman Chair in Colorectal Surgery.

Funding

This study was funded in part by the 2015 Society of American Gastrointestinal and Endoscopic Surgeons (SAGES) Career Development Award and the American Society of Colon and Rectal Surgeons (ASCRS) General Surgery Resident Research Initiation Grant #25 to Georgios Karagkounis.

Compliance with ethical standards

Disclosures

Georgios Karagkounis, Jennifer DeVecchio, Sylvain Ferrandon, and Matthew F. Kalady have no conflicts of interest or financial ties to disclose.

References

  1. 1.
    O’Connell MJ, Martenson JA, Wieand HS, Krook JE, Macdonald JS, Haller DG, Mayer RJ, Gunderson LL, Rich TA (1994) Improving adjuvant therapy for rectal cancer by combining protracted-infusion fluorouracil with radiation therapy after curative surgery. N Engl J Med 331(8):502–507. doi: 10.1056/nejm199408253310803 CrossRefPubMedGoogle Scholar
  2. 2.
    Kapiteijn E, Marijnen CA, Nagtegaal ID, Putter H, Steup WH, Wiggers T, Rutten HJ, Pahlman L, Glimelius B, van Krieken JH, Leer JW, van de Velde CJ (2001) Preoperative radiotherapy combined with total mesorectal excision for resectable rectal cancer. N Engl J Med 345(9):638–646. doi: 10.1056/NEJMoa010580 CrossRefPubMedGoogle Scholar
  3. 3.
    Sauer R, Becker H, Hohenberger W, Rodel C, Wittekind C, Fietkau R, Martus P, Tschmelitsch J, Hager E, Hess CF, Karstens JH, Liersch T, Schmidberger H, Raab R (2004) Preoperative versus postoperative chemoradiotherapy for rectal cancer. N Engl J Med 351(17):1731–1740. doi: 10.1056/NEJMoa040694 CrossRefPubMedGoogle Scholar
  4. 4.
    Sauer R, Liersch T, Merkel S, Fietkau R, Hohenberger W, Hess C, Becker H, Raab HR, Villanueva MT, Witzigmann H, Wittekind C, Beissbarth T, Rodel C (2012) Preoperative versus postoperative chemoradiotherapy for locally advanced rectal cancer: results of the German CAO/ARO/AIO-94 randomized phase III trial after a median follow-up of 11 years. J Clin Oncol 30(16):1926–1933. doi: 10.1200/jco.2011.40.1836 CrossRefPubMedGoogle Scholar
  5. 5.
    Rodel C, Martus P, Papadoupolos T, Fuzesi L, Klimpfinger M, Fietkau R, Liersch T, Hohenberger W, Raab R, Sauer R, Wittekind C (2005) Prognostic significance of tumor regression after preoperative chemoradiotherapy for rectal cancer. J Clin Oncol 23(34):8688–8696. doi: 10.1200/jco.2005.02.1329 CrossRefPubMedGoogle Scholar
  6. 6.
    Maas M, Nelemans PJ, Valentini V, Das P, Rodel C, Kuo LJ, Calvo FA, Garcia-Aguilar J, Glynne-Jones R, Haustermans K, Mohiuddin M, Pucciarelli S, Small W Jr, Suarez J, Theodoropoulos G, Biondo S, Beets-Tan RG, Beets GL (2010) Long-term outcome in patients with a pathological complete response after chemoradiation for rectal cancer: a pooled analysis of individual patient data. Lancet Oncol 11(9):835–844. doi: 10.1016/s1470-2045(10)70172-8 CrossRefPubMedGoogle Scholar
  7. 7.
    Mace AG, Pai RK, Stocchi L, Kalady MF (2015) American Joint Committee on Cancer and College of American Pathologists regression grade: a new prognostic factor in rectal cancer. Dis Colon Rectum 58(1):32–44. doi: 10.1097/dcr.0000000000000266 CrossRefPubMedGoogle Scholar
  8. 8.
    Agarwal B, Bhendwal S, Halmos B, Moss SF, Ramey WG, Holt PR (1999) Lovastatin augments apoptosis induced by chemotherapeutic agents in colon cancer cells. Clin Cancer Res 5(8):2223–2229PubMedGoogle Scholar
  9. 9.
    Chan KK, Oza AM, Siu LL (2003) The statins as anticancer agents. Clin Cancer Res 9(1):10–19PubMedGoogle Scholar
  10. 10.
    Graaf MR, Beiderbeck AB, Egberts AC, Richel DJ, Guchelaar HJ (2004) The risk of cancer in users of statins. J Clin Oncol 22(12):2388–2394. doi: 10.1200/jco.2004.02.027 CrossRefPubMedGoogle Scholar
  11. 11.
    Poynter JN, Gruber SB, Higgins PD, Almog R, Bonner JD, Rennert HS, Low M, Greenson JK, Rennert G (2005) Statins and the risk of colorectal cancer. N Engl J Med 352(21):2184–2192. doi: 10.1056/NEJMoa043792 CrossRefPubMedGoogle Scholar
  12. 12.
    Mace AG, Gantt GA, Skacel M, Pai R, Hammel JP, Kalady MF (2013) Statin therapy is associated with improved pathologic response to neoadjuvant chemoradiation in rectal cancer. Dis Colon Rectum 56(11):1217–1227. doi: 10.1097/DCR.0b013e3182a4b236 CrossRefPubMedGoogle Scholar
  13. 13.
    Katz MS, Minsky BD, Saltz LB, Riedel E, Chessin DB, Guillem JG (2005) Association of statin use with a pathologic complete response to neoadjuvant chemoradiation for rectal cancer. Int J Radiat Oncol Biol Phys 62(5):1363–1370. doi: 10.1016/j.ijrobp.2004.12.033 CrossRefPubMedGoogle Scholar
  14. 14.
    Konstantinopoulos PA, Karamouzis MV, Papavassiliou AG (2007) Post-translational modifications and regulation of the RAS superfamily of GTPases as anticancer targets. Nat Rev Drug Discov 6(7):541–555. doi: 10.1038/nrd2221 CrossRefPubMedGoogle Scholar
  15. 15.
    Huang EH, Johnson LA, Eaton K, Hynes MJ, Carpentino JE, Higgins PD (2010) Atorvastatin induces apoptosis in vitro and slows growth of tumor xenografts but not polyp formation in MIN mice. Dig Dis Sci 55(11):3086–3094. doi: 10.1007/s10620-010-1157-x CrossRefPubMedGoogle Scholar
  16. 16.
    Lee SJ, Lee I, Lee J, Park C, Kang WK (2014) Statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, potentiate the anti-angiogenic effects of bevacizumab by suppressing angiopoietin2, BiP, and Hsp90alpha in human colorectal cancer. Br J Cancer 111(3):497–505. doi: 10.1038/bjc.2014.283 CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Lee J, Lee I, Han B, Park JO, Jang J, Park C, Kang WK (2011) Effect of simvastatin on cetuximab resistance in human colorectal cancer with KRAS mutations. J Natl Cancer Inst 103(8):674–688. doi: 10.1093/jnci/djr070 CrossRefPubMedGoogle Scholar
  18. 18.
    Lee J, Hong YS, Hong JY, Han SW, Kim TW, Kang HJ, Kim TY, Kim KP, Kim SH, Do IG, Kim KM, Sohn I, Park SH, Park JO, Lim HY, Cho YB, Lee WY, Yun SH, Kim HC, Park YS, Kang WK (2014) Effect of simvastatin plus cetuximab/irinotecan for KRAS mutant colorectal cancer and predictive value of the RAS signature for treatment response to cetuximab. Invest New Drugs 32(3):535–541. doi: 10.1007/s10637-014-0065-x CrossRefPubMedGoogle Scholar
  19. 19.
    Kodach LL, Jacobs RJ, Voorneveld PW, Wildenberg ME, Verspaget HW, van Wezel T, Morreau H, Hommes DW, Peppelenbosch MP, van den Brink GR, Hardwick JC (2011) Statins augment the chemosensitivity of colorectal cancer cells inducing epigenetic reprogramming and reducing colorectal cancer cell ‘stemness’ via the bone morphogenetic protein pathway. Gut 60(11):1544–1553. doi: 10.1136/gut.2011.237495 CrossRefPubMedGoogle Scholar
  20. 20.
    Lacerda L, Reddy JP, Liu D, Larson R, Li L, Masuda H, Brewer T, Debeb BG, Xu W, Hortobagyi GN, Buchholz TA, Ueno NT, Woodward WA (2014) Simvastatin radiosensitizes differentiated and stem-like breast cancer cell lines and is associated with improved local control in inflammatory breast cancer patients treated with postmastectomy radiation. Stem Cells Transl Med 3(7):849–856. doi: 10.5966/sctm.2013-0204 CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Sanli T, Liu C, Rashid A, Hopmans SN, Tsiani E, Schultz C, Farrell T, Singh G, Wright J, Tsakiridis T (2011) Lovastatin sensitizes lung cancer cells to ionizing radiation: modulation of molecular pathways of radioresistance and tumor suppression. J Thorac Oncol 6(3):439–450. doi: 10.1097/JTO.0b013e3182049d8b CrossRefPubMedGoogle Scholar
  22. 22.
    Theodoropoulos G, Wise WE, Padmanabhan A, Kerner BA, Taylor CW, Aguilar PS, Khanduja KS (2002) T-level downstaging and complete pathologic response after preoperative chemoradiation for advanced rectal cancer result in decreased recurrence and improved disease-free survival. Dis Colon Rectum 45(7):895–903CrossRefPubMedGoogle Scholar
  23. 23.
    Demierre MF, Higgins PD, Gruber SB, Hawk E, Lippman SM (2005) Statins and cancer prevention. Nat Rev Cancer 5(12):930–942. doi: 10.1038/nrc1751 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Department of Colorectal SurgeryCleveland ClinicClevelandUSA
  2. 2.Department of Stem Cell Biology and Regenerative MedicineCleveland ClinicClevelandUSA

Personalised recommendations