Molecular targeting of prostate cancer during androgen ablation: Inhibition of CHES1/FOXN3

s and Presentations 1. Xiang, N., Ali, H.M., Davis, R.R., Liu, S.Y., Boucher, D.L., Gregg, J.P., Kung, H.J., and Tepper, C.G. Potential role of CHES1/FOXN3 as an anti-apoptotic regulator of prostate cancer response to androgen ablative and genotoxic therapies. Poster presented at: 15th Annual Cancer Research Symposium, October 28, 2010, UC Davis Cancer Center, Sacramento, CA. 2. Invited presentation, seminar entitled: “Integrative genomics approaches to understanding prostate cancer progression” November 12, 2010, National Tsing Hua University, Hsinchu, Taiwan, R.O.C. 3. Tepper, C.G., Xiang, N., Ali, H.M., Gregg, J.P., Yang, J.K., Boucher, D.L, Wee, C.B., Webb, J.M., Liu, S.Y., de Vere White, R.W., Gregg, J.P., and Kung, H.J. Molecular Targeting of Prostate Cancer during Androgen Ablation: Inhibition of CHES1/FOXN3. Poster presented at: Innovative Minds in Prostate Cancer Today (IMPaCT) 2011 Conference, March 9-12, 2011, Hilton Orlando, Orlando, FL. 4. Xiang, N., Purnell, S.M., Wee, C.B., Boucher, D.L., Shi, X.B., de Vere White, R.W., Gregg, J.P., Kung, H.J., and Tepper, C.G. Amino acid-mediated mTORC1 activation is a central integration point for androgen receptor and survival signaling in prostate cancer. Poster presented at: Keystone Symposium on "Cancer and Metabolism”, February 12-17, 2012, Fairmont Banff Springs, Banff, Alberta, Canada. Development of expression vectors and cell lines During the research period for this grant, we have generated a number of expression constructs and LNCaP sublines. These are listed below: 1. CHES1 RNA interference (RNAi, Ri) expression constructs: A. Constitutively expressed shRNA vectors: pSM2-CHES1-Ri-1, -2, and -3 (Fig. 9, 2010 Annual Report). B. Tetracycline/doxycycline-inducible shRNA expression constructs: 1) pTRIPZ-CHES1-Ri-1, -3, and -5 (Fig. 11, 2010 Annual Report). 2) Tetracycline/doxycycline-inducible CHES1 expression construct: pRev-TRE-HACHES1. 3) Constitutive CHES1/FOXN3 expression construct with FLAG-epitope tag: pcDNA3.1-FLAG-CHES1. 2. Stable LNCaP and CWR22Pc cell lines with constitutive expression of CHES1 shRNAs: A. LNCaP-CHES1-Ri-1 through -9 (Fig. 9, 2010 Annual Report) and LNCaP-pSM2 vector control. B. CWR22Pc-CHES1-Ri-1/3/7 and CWR22Pc-pSM2 vector control. 3. LNCaP cells with Tet/Dox-inducible CHES1-targeting shRNA expression: LNCaP-tetCHES1-Ri-1, -3, and -5 (Fig. 11, 2010 Annual Report) and LNCaP-pTRIPZ empty vector control. 4. CHES1/FOXN3 expression constructs: A. Tet/Dox-inducible CHES1 expression construct: pLVX-Tight-Puro-FLAG-CHES1. 5. Stable LNCaP sublines with Tet/Dox-inducible CHES1/FOXN3 expression (LNCaP-tetFLAG-CHES1) and corresponding vector-control cell line, LNCaP-tet-pLVX. 6. Stable LNCaP and 22Rv1 cell lines with constitutive expression of epitope-tagged mTOR, raptor, RagB-wt, RagB-GDP, RagB-GTP, RagD-wt, RagD-GDP, RagD-GTP. 7. Luciferase reporter constructs: A. CHES1 Firefly luciferase reporter construct: pGL3B-CHES1-RR1/3.5 B. BNIP3 Firefly luciferase reporter construct: pGL3B-BNIP3(4.0)