Abstract P4-07-09: MYB is involved in the DNA damage response in human ER+ breast cancer cells
Authors: R-M Yang, E Dray, P Mitra, TJ Gonda
Journal: Cancer Research (2016)

Abstract Aims: Over 70% of human breast cancer cells are oestrogen receptor positive (ER+) and express MYB. MYB expression is necessary for the proliferation of ER+ breast cancer cells in vitro and for tumour development in vivo. Our previous studies found that shRNA-mediated MYB knock-down greatly sensitised breast cancer cells to chemically-induced apoptosis by down-regulating the BCL2 (a MYB target gene) (Drabsch et al., 2010). Furthermore, several published studies (Taha et al., 2004; Thomadaki et al., 2006) indicated that actinomycin D and etoposide treatment could induce DNA damage in human ER+ breast cancer (MCF-7) cells. Moreover, silencing MYB increased DNA damage-induced cell death in castration resistant prostate cancer cells by down-regulating DNA damage response genes (Li et al., 2014). However, there is very little information on MYB function in the DNA damage response of ER+ breast cancer cells. Therefore, the aim of this study was to investigate whether silencing MYB affected the DNA damage repair and resulting in cell death in MCF-7 cells. Methods: To achieve down-regulation of MYB expression in human ER+ breast cancer, we used doxycycline inducible shRNA lentiviral vectors (pLV711 (Drabsch et al., 2007; Brown et al., 2010)) in human MCF-7 breast cancer cells. We used PI staining plus FACS analysis for cell death assessment. We also used γ-H2AX protein expression and γ-H2AX foci counting to assess DNA damage. Results: We found that silencing MYB alone did not result in cell death, as reported previously (Drabsch et al., 2010). However, silencing MYB significantly increased actinomycin D-induced cell death. This similar result was also found on etoposide-induced cell death. Furthermore, we found that silencing MYB significantly increased actinomycin D or etoposide-induced γ-H2AX expression. Moreover, anti-apoptotic BCL2 expression, measured by western blotting, was dramatically reduced after the combination of MYB knock down and actinomycin D or etoposide, compared to wild type and nonsilencing controls. Conclusions: Silencing MYB significantly increased DNA damage-induced cell death and D???damage. This may result from down-regulation of BCL2 expression, an effect on DNA damage response genes, or both. This requires further investigation. For example, Rad51 and γ-H2AX colocalization and 53BP1 expression in response to DNA damage will be presented. References: Taha et al., 2004, J Biol Chem, 279, 20546-46. Thomadaki et al., 2006, Biol. Chem., 387, 1081-1086. Drabsch et al., 2007, Proc Natl Acad Sci U S A, 104, 13762-7. Drabsch et al., 2010, Breast Cancer Res, 12:R55. Brown et al., 2010, Hum Gene Ther, 21(8),1005-17. Li et al., 2014, Science Signaling, 7(326). Citation Format: Yang R-M, Dray E, Mitra P, Gonda TJ. MYB is involved in the DNA damage response in human ER+ breast cancer cells. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-07-09.