Antiestrogen-resistant and triple-negative breast tumors pose a serious clinical challenge because of limited treatment options. We assessed global gene expression changes in antiestrogen-sensitive compared with antiestrogen-resistant (two tamoxifen resistant and two fulvestrant resistant) MCF-7 breast cancer cell lines. The branched-chain amino acid transaminase 1 (BCAT1), which catalyzes the first step in the breakdown of branched-chain amino acids, was among the most upregulated transcripts in antiestrogen-resistant cells. Elevated BCAT1 expression was confirmed in relapsed tamoxifen-resistant breast tumor specimens. High intratumoral BCAT1 levels were associated with a reduced relapse-free survival in adjuvant tamoxifen-treated patients and overall survival in unselected patients. On a tissue microarray (n=1421), BCAT1 expression was detectable in 58% of unselected primary breast carcinomas and linked to a higher Ki-67 proliferation index, as well as histological grade. Interestingly, BCAT1 was predominantly expressed in estrogen receptor-α-negative/human epidermal growth factor receptor-2-positive (ERα-negative/HER-2-positive) and triple-negative breast cancers in independent patient cohorts. The inverse relationship between BCAT1 and ERα was corroborated in various breast cancer cell lines and pharmacological long-term depletion of ERα induced BCAT1 expression in vitro. Mechanistically, BCAT1 indirectly controlled expression of the cell cycle inhibitor p27Kip1 thereby affecting pRB. Correspondingly, phenotypic analyses using a lentiviral-mediated BCAT1 short hairpin RNA knockdown revealed that BCAT1 sustains proliferation in addition to migration and invasion and that its overexpression enhanced the capacity of antiestrogen-sensitive cells to grow in the presence of antiestrogens. Importantly, silencing of BCAT1 in an orthotopic triple-negative xenograft model resulted in a massive reduction of tumor volume in vivo, supporting our findings that BCAT1 is necessary for the growth of hormone-independent breast tumors.
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We highly appreciate the excellent technical assistance of Karin Pfleger and Achim Stephan. Further, we thank Nicolas Hafner and Yashna Paul for experimental and computational support and appreciate the help and constructive discussions from Martina Seiffert, and Michael Fletcher. Moreover, we thank the patients, pathologists and gynecologists associated with the PATH Biobank (http://path-biobank.org/index.php/en/) for their support. This work was supported by the Strategic Alliance of Bayer Healthcare and the German Cancer Research Center (DKFZ). VT was supported by a scholarship from the Helmholtz-Graduate School for Cancer Research.
VT, MH, UD, AS, SH, AEL, MZ, BR, PL and MT conceived the project and designed the experiments. VT, MS, PS and MT performed in vitro experiments. YW and WW conducted MassARRAY analyses. KS and JGO conducted mass spectrometry analyses. TA, NM, RB, HPS, AS and GS provided and evaluated tumor specimens and clinical data. VT, RS, NM, HPS, AS, RB and MZ generated and analyzed clinical data. MH, MZ and RS performed computational analyses. MS prepared lentiviral particles with help of PW, SK and HH-S conducted animal experiments. PS and MK prepared xenograft sections with the help of NK. VT, UD, SH, MZ, BR, PL and MT analyzed data and wrote the manuscript with the help of other authors.
Stefan Kaulfuss and Holger Hess-Stumpp are employees of Bayer Pharma AG. However, no respective chemical compounds were used in the manuscript and the collaboration was for scientific purposes. The remaining authors declare no conflict of interest.
Supplementary Information accompanies this paper on the Oncogene website
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Thewes, V., Simon, R., Hlevnjak, M. et al. The branched-chain amino acid transaminase 1 sustains growth of antiestrogen-resistant and ERα-negative breast cancer. Oncogene 36, 4124–4134 (2017). https://doi.org/10.1038/onc.2017.32
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