癸 巳 年

1. 辞旧迎新蛇年大吉 2013 癸巳年 Androgen Receptor Regulates a Distinct Transcription Program in Androgen-Independent Prostate Cancer Reporter:zhaopingLiao Instructor:zhihuaTao Happy New Year

2. 2013 癸巳年 一.基本概念 顺反组(cistrome)是由“Dana-Farber癌症研究所”与哈佛医学院的科学家提出的遗传学术语， 用于定义一个反式（trans）调控因子在基因组（genome）范围内的作用对象(一组顺式(cis) 作用元素)。一些技术，例如免疫共沉淀与基因芯片结合的技术(ChIP-on-chip)，已经被广泛 的应用于发现转录因子以及其他染色质相关因子的顺反组。 顺式作用元件 (cis-actingelement)存在于基因旁侧序列中能影响基因表达的序列。顺式作用 元件包括启动子、增强子、调控序列和可诱导元件等，它们的作用是参与基因表达的调控。 顺式作用元件本身不编码任何蛋白质，仅仅提供一个作用位点，要与反式作用因子相互作 用而起作用。 反式调节因子（ trans-acting factor ）是指直接或能间接地识别或结合在各类顺式作用元件 核心序列上参与调控靶基因转录效率的蛋白质。有时也称转录因子。 大多数真核转录调节 因子由某一基因表达后，可通过另一基因的特异的顺式作用元件相互作用，从而激活另一 基因的转录。这种调节蛋白称反式作用因子。 癸巳年

3. 2013 癸巳年 ChIP-on-chip M-phase cell-cycle genes UBE2C（H3K4、FoxA1） inactivated M-phase checkpoint CELL CYCLE AND AIPC? 癸巳年

4. 2013 癸巳年 二.Role of AR in ADPC and AIPC? 1.Decreasing level of AR protein expression reduces both ADPC and AIPC growth in model systems; 2.Decreased ADPC growth after AR silencing and/or androgen deprivation primarily involves a block of the G1/S cell cycle transition through AR-dependent regulation of cyclin D1,p21 and P27; 3.Experiment support: L总 F总 L 核F核L胞F胞 AR GAPDH 癸巳年

5. 2013 癸巳年 三.Research subjects LNCaP-abl (abl) LNCaP C4-2B tissue material Culig, Z., Hoffmann, J., Erdel, M., Eder, I.E., Hobisch, A., Hittmair, A., Bartsch,G.,Utermann,G.,Schneider,M.R.,Parczyk,K., et al. (1999).Switch from antagonist to agonist of the androgen receptor bicalutamide is associated with prostate tumour progression in a new model system. Br. J. Cancer 81, 242–251. 癸巳年

6. 2013 癸巳年 四.results • M-phase cell-cycle genes are upregulated genes both in a cell culture model of • AIPC and in clinical AIPC samples • ①using affymetrix U133 plus 2.0 expression arrays to identify genes in abl in the • Absence of DHT and LNCaP cell in the absence and presence of DHT for 4 h combined • With Gene Ontology(GO) analysis of upregulated transceipts reveal “cell-cycle ”and • “M-phase”as top enriched GO processes; • ②cell –cycle genes are also enriched in upregulated genes from cases of AIPC • Compared with ADPC cases; • Conclusin:expression of cell-cycle regulatory genes,primarily M-phase genes are • enriched In AIPC and may promote AIPC growth. 癸巳年

7. 2013 癸巳年 2. AR regulate M-phase Cell-Cycle Genes to promote AIPC Growth “Cellular lipid metabolism” “Positive regulation of cellular process” 癸巳年

8. 2013 癸巳年 3.Preferential AR binding to the M-phase genes leads to higher expression in AIPC ChIP-on-chip 癸巳年

9. 2013 癸巳年 4.Selective active epigenetic marks and collaborating transcription factors at M-phase Gene enhancers lead to increased AR occupancy at these sites in AIPC ① Ubiquitin-conjugating enzyme E2C(UBE2C ) 3C-qPCR to test the putative enhancers communicate with the UBE2C promotor 癸巳年

10. 2013 癸巳年 ②coregulator---FoxA1 may play a more important role in AIPC than in ADPC ③H3K4 methylation(KDM1—demethylase) 癸巳年

11. 2013 癸巳年 ④relationship among H3K4 methylation、FoxA1 binding and AR binding 癸巳年

12. 2013 癸巳年 5. Higher levels of H3K4 metylation and FoxA1 binding at the UBE2C enhancers leads to overexpression of UBE2C protein in AIPC cases C4-2B cell lines and clinical cases of AIPC AR、FoxA1、H3K4me1、 H3K4me2 ChIP from clinical tissues also proved enhanced AR transcription complex occupancy at the UBE2C enhancers 癸巳年

13. 2013 癸巳年 6. Functional role of UBE2C in AIPC growth UBE2C protein Half-life of 6h UBE2C inactivating M-phase checkpoint and silence of it cause G2/M accumulation; siAR cause G1/S block in LNCaP but leads to G2/S block in abl cells 癸巳年

14. 2013 癸巳年 Silence of UBE2C and AR leads the accumulation of S、G2/M Cyclin A is not detected during early G1 ,accumulates an the end of G1 and id essential for G1/S transition UBE2C silence results in a shortened G1 phase in LNCaP but not in abl cells. UBE2C has a general role in accelerating M-phase transition in solid tumors,silencing of UBE2C significantly decrease AIPC growth by arresting G2/M and S phases, providing a potential new Target for therapeutic intervention 癸巳年