Advertisement

Tumor Biology

, Volume 37, Issue 4, pp 4323–4330 | Cite as

SAHA, an HDAC inhibitor, overcomes erlotinib resistance in human pancreatic cancer cells by modulating E-cadherin

  • Seong Joon Park
  • Seung-Mi Kim
  • Jai-Hee Moon
  • Jeong Hee Kim
  • Jae-Sik Shin
  • Seung-Woo Hong
  • Yu Jin Shin
  • Dae-Hee Lee
  • Eun Young Lee
  • Ih-Yeon Hwang
  • Jeong Eun Kim
  • Kyu-pyo Kim
  • Yong Sang Hong
  • Won–Keun Lee
  • Eun Kyung Choi
  • Jung Shin Lee
  • Dong-Hoon Jin
  • Tae Won Kim
Original Article

Abstract

Pancreatic cancer is one of the most lethal cancers and remains a major unsolved health problem. Less than 20 % of patients are surgical candidates, and the median survival for non-resected patients is approximately 3 to 4 months. Despite the existence of many conventional cancer therapies, few targeted therapies have been developed for pancreatic cancer. Combination therapy using erlotinib and gemcitabine is an approved standard chemotherapy for advanced pancreatic cancer, but it has marginal therapeutic benefit. To try to improve the therapeutic outlook, we studied the efficacy of another combination treatment and the relevance to E-cadherin in human pancreatic cancer cells. We treated two human pancreatic cancer cell lines with the histone deacetylase inhibitor (HDACi) SAHA. Interestingly, in these Panc-1 and Capan1 cells, we observed that the expression levels of E-cadherin and phosphorylated EGFR were gradually upregulated after treatment with SAHA. Furthermore, these cells underwent induced cell death after exposure to the combination treatment of SAHA and erlotinib. In Panc-1 cells, overexpression of E-cadherin activated the phosphorylation of EGFR and increased the cell sensitivity to erlotinib. In Capan1 cells, knocking down E-cadherin decreased the expression of phosphorylated EGFR, and these cells did not respond to erlotinib. Therefore, we demonstrated the efficacy of the combined treatment with SAHA and erlotinib in human pancreatic cancer cells, and we determined that the increased efficacy was due, at least in part, to the effects of SAHA on the expression of E-cadherin. Our studies suggest that E-cadherin may be a potent biomarker for pancreatic cancer.

Keywords

SAHA Erlotinib E-cadherin Human pancreatic cancer cells Cell death 

Abbreviations

HDACis

Histone deacetylase inhibitors

EGFR-TKI

EGFR tyrosine kinase inhibitor

Notes

Acknowledgments

This work was supported by grants from the Korea Health 21 R&D project, the Ministry of Health and Welfare and Family Affairs, Republic of Korea (HI06C0868), the Basic Science Research Program through the National Research Foundation of Korea (NRF), which was funded by the Ministry of Education, Science and Technology (2013R1A1A2013233), and the Asan Institute for Life Sciences, Seoul, Republic of Korea (2014-231).

Compliance with ethical standards

Conflicts of interest

None

Supplementary material

13277_2015_4216_MOESM1_ESM.eps (1.3 mb)
Supplementary Fig. 1 The HDAC inhibitor (SAHA) decreased cell viability in the human human pancreatic cancer cells (Panc08.13 and BxPC3). (a) Panc08.13 and (b) BxPC3 cells were treated with the indicated doses of SAHA for 48 hrs, and then cell death was determined using a trypan blue exclusion assay. Cells were harvested and analyzed by immunoblot using antibodies against E-cadherin, phospho-EGFR, and γ-tubulin. γ-tubulin was used as a loading control. The values are presented as the means ± SDs from three separate experiments that were performed in triplicate. *P<0.05, **P<0.01. (EPS 1365 kb)
13277_2015_4216_MOESM2_ESM.eps (1 mb)
Supplementary Fig. 2 SAHA decreased cell viability in human pancreatic cancer cells, PL45. PL45 cells were treated with the indicated doses of SAHA for 48 hrs and then cell death was measured by trypan blue exclusion assay. Cells were harvested and analyzed by immunoblot using antibodies against E-cadherin, phospho-EGFR, and γ-tubulin. γ-tubulin was used as a loading control. The values are presented as the means ± SDs from three separate experiments that were performed in triplicate. *P<0.05, **P<0.01. (EPS 1055 kb)
13277_2015_4216_MOESM3_ESM.eps (882 kb)
Supplementary Fig. 3 SAHA regulates transcriptional of E-cadherin. Cells were treated with SAHA for 48 hrs. mRNA levels of ZEB1, E47, and E-cadherin were detected by RT-PCR analysis. GAPDH was used as a loading control. (EPS 881 kb)
13277_2015_4216_MOESM4_ESM.eps (1.2 mb)
Supplementary Fig. 4 SAHA altered the cell morphology and regulated the expression of EMT markers in Panc-1 and Capan1 cells. After treatment with SAHA, the changes in these cells were observed by phase contrast light microscopy (left panel). The cells were harvested and analyzed for the expression of EMT-related genes by RT-PCR (right panel). (EPS 1208 kb)
13277_2015_4216_MOESM5_ESM.eps (1.3 mb)
Supplementary Fig. 5 Combined treatment with an EMT inhibitor (SB431542) and erlotinib induced cell death in Panc-1 and Capan1 cells. These cells showed increased cell death rates following treatment with SB431542 and erlotinib (left panel). The cells were harvested and analyzed by western blot using antibodies against E-cadherin, phospho-EGFR, and γ-tubulin. γ-tubulin was used as a loading control. The values are presented as the mean ± SD from three separate experiments that were performed in triplicate. *P<0.05, **P<0.01. (EPS 1341 kb)

References

  1. 1.
    Rossi ML, Rehman AA, Gondi CS. Therapeutic options for the management of pancreatic cancer. World J Gastroenterol. 2014;20(32):11142–59. doi: 10.3748/wjg.v20.i32.11142.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Becker AE, Hernandez YG, Frucht H, Lucas AL. Pancreatic ductal adenocarcinoma: risk factors, screening, and early detection. World J Gastroenterol. 2014;20(32):11182–98. doi: 10.3748/wjg.v20.i32.11182.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Teague A, Lim KH, Wang-Gillam A. Advanced pancreatic adenocarcinoma: a review of current treatment strategies and developing therapies. Ther Adv Med Oncol. 2015;7(2):68–84. doi: 10.1177/1758834014564775.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Kleeff J, Reiser C, Hinz U, Bachmann J, Debus J, Jaeger D, et al. Surgery for recurrent pancreatic ductal adenocarcinoma. Ann Surg. 2007;245(4):566–72. doi: 10.1097/01.sla.0000245845.06772.7d.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Moore MJ, Goldstein D, Hamm J, Figer A, Hecht JR, Gallinger S, et al. Erlotinib plus gemcitabine compared with gemcitabine alone in patients with advanced pancreatic cancer: a phase III trial of the National Cancer Institute of Canada Clinical Trials Group. J Clin Oncol. 2007;25(15):1960–6. doi: 10.1200/JCO.2006.07.9525.CrossRefPubMedGoogle Scholar
  6. 6.
    Dokmanovic M, Clarke C, Marks PA. Histone deacetylase inhibitors: overview and perspectives. Mol Cancer Res. 2007;5(10):981–9. doi: 10.1158/1541-7786.MCR-07-0324.CrossRefPubMedGoogle Scholar
  7. 7.
    Takai N, Desmond JC, Kumagai T, Gui D, Said JW, Whittaker S, et al. Histone deacetylase inhibitors have a profound antigrowth activity in endometrial cancer cells. Clin Cancer Res. 2004;10(3):1141–9.CrossRefPubMedGoogle Scholar
  8. 8.
    Marks PA. Discovery and development of SAHA as an anticancer agent. Oncogene. 2007;26(9):1351–6. doi: 10.1038/sj.onc.1210204.CrossRefPubMedGoogle Scholar
  9. 9.
    Arnold NB, Arkus N, Gunn J, Korc M. The histone deacetylase inhibitor suberoylanilide hydroxamic acid induces growth inhibition and enhances gemcitabine-induced cell death in pancreatic cancer. Clin Cancer Res. 2007;13(1):18–26. doi: 10.1158/1078-0432.CCR-06-0914.CrossRefPubMedGoogle Scholar
  10. 10.
    Lange F, Rateitschak K, Kossow C, Wolkenhauer O, Jaster R. Insights into erlotinib action in pancreatic cancer cells using a combined experimental and mathematical approach. World J Gastroenterol. 2012;18(43):6226–34. doi: 10.3748/wjg.v18.i43.6226.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Witta SE, Gemmill RM, Hirsch FR, Coldren CD, Hedman K, Ravdel L, et al. Restoring E-cadherin expression increases sensitivity to epidermal growth factor receptor inhibitors in lung cancer cell lines. Cancer Res. 2006;66(2):944–50. doi: 10.1158/0008-5472.CAN-05-1988.CrossRefPubMedGoogle Scholar
  12. 12.
    Black PC, Brown GA, Inamoto T, Shrader M, Arora A, Siefker-Radtke AO, et al. Sensitivity to epidermal growth factor receptor inhibitor requires E-cadherin expression in urothelial carcinoma cells. Clin Cancer Res. 2008;14(5):1478–86. doi: 10.1158/1078-0432.CCR-07-1593.CrossRefPubMedGoogle Scholar
  13. 13.
    Qian X, Karpova T, Sheppard AM, McNally J, Lowy DR. E-cadherin-mediated adhesion inhibits ligand-dependent activation of diverse receptor tyrosine kinases. EMBO J. 2004;23(8):1739–48. doi: 10.1038/sj.emboj.7600136.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Chen MC, Chen CH, Wang JC, Tsai AC, Liou JP, Pan SL, et al. The HDAC inhibitor, MPT0E028, enhances erlotinib-induced cell death in EGFR-TKI-resistant NSCLC cells. Cell Death Dis. 2013;4:e810. doi: 10.1038/cddis.2013.330.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Huber O, Kemler R, Langosch D. Mutations affecting transmembrane segment interactions impair adhesiveness of E-cadherin. J Cell Sci. 1999;112(Pt 23):4415–23.PubMedGoogle Scholar
  16. 16.
    Lewis-Tuffin LJ, Rodriguez F, Giannini C, Scheithauer B, Necela BM, Sarkaria JN, et al. Misregulated E-cadherin expression associated with an aggressive brain tumor phenotype. PLoS One. 2010;5(10):e13665. doi: 10.1371/journal.pone.0013665.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Wang Y, Zhou BP. Epithelial-mesenchymal transition—a hallmark of breast cancer metastasis. Cancer Hallm. 2013;1(1):38–49. doi: 10.1166/ch.2013.1004.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Pecina-Slaus N. Tumor suppressor gene E-cadherin and its role in normal and malignant cells. Cancer Cell Int. 2003;3(1):17. doi: 10.1186/1475-2867-3-17.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Aghdassi A, Sendler M, Guenther A, Mayerle J, Behn CO, Heidecke CD, et al. Recruitment of histone deacetylases HDAC1 and HDAC2 by the transcriptional repressor ZEB1 downregulates E-cadherin expression in pancreatic cancer. Gut. 2012;61(3):439–48. doi: 10.1136/gutjnl-2011-300060.CrossRefPubMedGoogle Scholar
  20. 20.
    Halder SK, Beauchamp RD, Datta PK. A specific inhibitor of TGF-beta receptor kinase, SB-431542, as a potent antitumor agent for human cancers. Neoplasia. 2005;7(5):509–21.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Seong Joon Park
    • 1
    • 2
  • Seung-Mi Kim
    • 1
    • 2
    • 3
  • Jai-Hee Moon
    • 1
    • 2
  • Jeong Hee Kim
    • 1
    • 2
  • Jae-Sik Shin
    • 1
    • 2
  • Seung-Woo Hong
    • 1
    • 2
  • Yu Jin Shin
    • 1
    • 2
  • Dae-Hee Lee
    • 1
    • 2
  • Eun Young Lee
    • 1
    • 2
  • Ih-Yeon Hwang
    • 1
    • 2
  • Jeong Eun Kim
    • 1
    • 2
  • Kyu-pyo Kim
    • 1
    • 2
  • Yong Sang Hong
    • 1
    • 2
  • Won–Keun Lee
    • 3
  • Eun Kyung Choi
    • 1
    • 4
  • Jung Shin Lee
    • 1
    • 2
  • Dong-Hoon Jin
    • 1
    • 2
    • 5
  • Tae Won Kim
    • 1
    • 2
  1. 1.Innovative Cancer ResearchASAN Institute for Life Science, Asan Medical CenterSeoulSouth Korea
  2. 2.Department of OncologyUniversity of Ulsan College of Medicine, Asan Medical CenterSeoulSouth Korea
  3. 3.Department of Biosciences and BioinformaticsMyongji UniversityYongin-siSouth Korea
  4. 4.Department of Radiation OncologyUniversity of Ulsan College of Medicine, Asan Medical CenterSeoulSouth Korea
  5. 5.Department of Convergence MedicineUniversity of Ulsan College of Medicine, Asan Medical CenterSeoulSouth Korea

Personalised recommendations