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Role of PTEN promoter methylation in tamoxifen-resistant breast cancer cells

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Abstract

Tamoxifen (TAM) resistance is a serious clinical problem in the treatment of breast cancer. Here, we found that S-adenosylmethionine (SAM) and DNA methyltransferase1 (DNMT1) expression are up-regulated in TAM-resistant breast cancer (TAMR-MCF-7) cells. We further focused on whether increased SAM with DNMT1 overexpression in TAMR-MCF-7 cells lead to aberrant methylation of the PTEN gene promoter and its therapeutic potential. Methylation-specific PCR analyses revealed that two sites within the PTEN promoters were methylated in TAMR-MCF-7 cells, which resulted in down-regulation of PTEN expression and increase in Akt phosphorylation. Both the loss of PTEN expression and the increased Akt phosphorylation in TAMR-MCF-7 cells were completely reversed by 5-aza-2′-deoxycytidine (5-Aza), a DNMT inhibitor. 5-Aza inhibited the basal cell proliferation rate of TAMR-MCF-7 cells and intraperitoneal injection of 5-Aza significantly suppressed TAMR-MCF-7 tumor growth in a xenograft study. Immunohistochemistry showed that PTEN expression in TAM-resistant human breast cancer tissues was lower than in TAM-responsive cases. These results suggest that methylation of the PTEN promoter related to both SAM increase and DNMT1 activation contributes to persistent Akt activation and are potential therapeutic targets for reversing TAM resistance in breast cancer.

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Abbreviations

5-Aza:

5-Aza-2′-deoxycytidine

BRCA1 :

Breast cancer 1

DMEM:

Dulbecco’s modified Eagle’s medium

DNMT:

DNA methyltranferase

ERα :

Estrogen receptor α

GSTP1 :

Glutathione S-transferase P

MAT:

Methionine adenosyltransferase

PCNA:

Proliferating cell nuclear antigen

PI3K:

Phosphoinositide 3-kinases

PTEN:

Phosphatase and tensin homolog

P16INK4A :

Cyclin-dependent kinase inhibitor 2A

SAM:

S-adenosylmethionine

TAM:

Tamoxifen

TIMP-3:

Metalloproteinase inhibitor 3

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

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Acknowldegment

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (Keon Wook Kang, No. 2009-0070587).

Conflicts of interest

All authors declare no conflicts of interest.

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Authors and Affiliations

  1. BK21 Project Team, College of Pharmacy, Chosun University, Gwangju, 501-759, South Korea

    Nguyen Thi Thuy Phuong & Keon Wook Kang

  2. College of Pharmacy, Chungnam National University, Daejeon, South Korea

    Sang Kyum Kim

  3. Department of Pathology, College of Medicine, Chosun University, Gwangju, 501-759, South Korea

    Sung Chul Lim

  4. College of Pharmacy, Pusan National University, Busan, 609-735, South Korea

    Hyung Sik Kim & Tae Hyung Kim

  5. College of Pharmacy, Chonnam National University, Gwangju, 500-757, South Korea

    Kwang Yeol Lee

  6. Department of Pathology, Research Center for Oral Disease Regulation of the Aged, School of Dentistry, Chosun University, Gwangju, 500-759, South Korea

    Sang-Gun Ahn & Jung-Hoon Yoon

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  1. Nguyen Thi Thuy Phuong
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  2. Sang Kyum Kim
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Correspondence to Keon Wook Kang.

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N. T. T. Phuong and S. K. Kim equally contributed to this work.

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Phuong, N.T.T., Kim, S.K., Lim, S.C. et al. Role of PTEN promoter methylation in tamoxifen-resistant breast cancer cells. Breast Cancer Res Treat 130, 73–83 (2011). https://doi.org/10.1007/s10549-010-1304-2

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  • DOI: https://doi.org/10.1007/s10549-010-1304-2

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