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

Tumor Biology

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.

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Abbreviations

HDACis:

Histone deacetylase inhibitors

EGFR-TKI:

EGFR tyrosine kinase inhibitor

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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).

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Correspondence to Dong-Hoon Jin or Tae Won Kim.

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Seong Joon Park, Seung-Mi Kim and Jai-Hee Moon contributed equally to this work.

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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)

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)

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)

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)

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)

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Park, S.J., Kim, SM., Moon, JH. et al. SAHA, an HDAC inhibitor, overcomes erlotinib resistance in human pancreatic cancer cells by modulating E-cadherin. Tumor Biol. 37, 4323–4330 (2016). https://doi.org/10.1007/s13277-015-4216-2

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  • DOI: https://doi.org/10.1007/s13277-015-4216-2

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