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Tumor Biology

, Volume 37, Issue 4, pp 4743–4753 | Cite as

SIRT1 induces tumor invasion by targeting epithelial mesenchymal transition-related pathway and is a prognostic marker in triple negative breast cancer

  • Min-Sun Jin
  • Chang Lim Hyun
  • In Ae Park
  • Ji Young Kim
  • Yul Ri Chung
  • Seock-Ah Im
  • Kyung-Hun Lee
  • Hyeong-Gon Moon
  • Han Suk Ryu
Original Article

Abstract

Absence of therapeutic targets poses a critical hurdle in improving prognosis for patients with triple negative breast cancer (TNBC). We evaluated interaction between SIRT1 and epithelial mesenchymal transition (EMT)-associated proteins as well as the role of combined protein expression as a predictor of lymph node metastasis and clinical outcome in TNBC through in vivo and vitro studies. Three hundred nineteen patients diagnosed with TNBC were chosen, immunohistochemical staining for SIRT1 and EMT-related markers’ expression was performed on tissue microarrays, and in vitro experiments with each of the three human TNBC cell lines were carried out. The cohort was reclassified according to the use of adjuvant chemotherapy, tumor size, and AJCC stage to analyze the prognostic role of SIRT1 and EMT-related proteins’ expression considering different therapeutic modalities and AJCC stages. Combination of four proteins including SIRT1 and three EMT-related proteins was revealed to be a statistically significant independent predictor of lymph node metastasis in the tumor size cohort as well as in the total patient population. Upon Cox regression analysis, increased expression level of the combined proteins correlated with decreased disease-free survival in the total patients as well as those who received adjuvant chemotherapy and those who had early stage breast cancer. In additional in vitro experiments, inhibition of SIRT1 expression with small interfering RNA (siRNA) suppressed tumor invasion in three different TNBC cell lines, and altered expression levels of EMT-related proteins following SIRT1 gene inhibition were identified on western blotting and fluorescence activated cell sorting (FACS) analysis; on the other hand, no change in expression levels of the cell cycle-related factors was observed. Our analysis showed the potential role of SIRT1 in association with EMT-related factors on tumor invasion, metastasis, and disease-free survival in TNBC, SIRT1, and associated EMT-related markers may offer a new prognostic indicator as well as a novel therapeutic candidate.

Keywords

Triple negative breast cancer SIRT1 Epithelial mesenchymal transition Metastasis Invasion Disease-free survival 

Notes

Acknowledgments

This study was supported by Grant No. 03-2014-0380 from Seoul National University Hospital Research Fund and by a research grant from Jeju National University Hospital in 2013–15.

Compliance with ethical standards

Conflicts of interest

None

Supplementary material

13277_2015_4231_MOESM1_ESM.doc (42 kb)
Supplementary Table 1 List of antibodies used in this study (DOC 42 kb)
13277_2015_4231_MOESM2_ESM.doc (42 kb)
Supplementary Table 2 Comparisons of clinicopathologic variables predicting lymph node metastasis using Pearson chi square test in 319 TNBC patients (DOC 42 kb)
13277_2015_4231_MOESM3_ESM.doc (32 kb)
Supplementary Table 3 Correlation coefficients of four proteins expression (DOC 31 kb)
13277_2015_4231_MOESM4_ESM.doc (46 kb)
Supplementary Table 4 Kaplan-Meier survival analysis for disease-free and overall survival (DOC 45 kb)
13277_2015_4231_MOESM5_ESM.doc (50 kb)
Supplementary Table 5 Kaplan-Meier survival analysis for disease-free survival in stratified TNBC cohort based on adjuvant chemotherapy and AJCC stage (DOC 50 kb)
13277_2015_4231_MOESM6_ESM.doc (51 kb)
Supplementary Table 6 Kaplan-Meier survival analysis for overall survival in stratified TNBC cohort based on adjuvant chemotherapy and AJCC stage (DOC 51 kb)
13277_2015_4231_Fig5_ESM.gif (26 kb)
Supplementary Fig. 1

ROC curve for triple negative breast cancer. (GIF 25 kb)

13277_2015_4231_MOESM7_ESM.tif (2.1 mb)
High resolution image (TIFF 2169 kb)

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Min-Sun Jin
    • 1
  • Chang Lim Hyun
    • 2
  • In Ae Park
    • 3
  • Ji Young Kim
    • 3
  • Yul Ri Chung
    • 3
  • Seock-Ah Im
    • 4
  • Kyung-Hun Lee
    • 4
  • Hyeong-Gon Moon
    • 5
  • Han Suk Ryu
    • 3
  1. 1.Department of PathologyBuchen St. Mary’s Hospital, Catholic universityGyeonggi-doSouth Korea
  2. 2.Department of PathologyJeju National University HospitalJejuSouth Korea
  3. 3.Department of PathologySeoul National University Hospital, Seoul National University College of MedicineSeoulSouth Korea
  4. 4.Department of Internal MedicineSeoul National University HospitalSeoulSouth Korea
  5. 5.Department of SurgerySeoul National University HospitalSeoulSouth Korea

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