Zimmer Brenna M, Howell Michelle E, Wei Qin, Ma Linlin, Romsdahl Trevor, Loughman Eileen G, Markham Jennifer E, Seravalli Javier, Barycki Joseph J, Simpson Melanie A
Department of Biochemistry, University of Nebraska, 1901 Vine Street, Lincoln, NE, 68588-0664, USA.
Horm Cancer. 2016 Aug;7(4):260-71. doi: 10.1007/s12672-016-0268-z. Epub 2016 Jun 15.
Prostate epithelial cells control the potency and availability of androgen hormones in part by inactivation and elimination. UDP-glucose dehydrogenase (UGDH) catalyzes the NAD(+)-dependent oxidation of UDP-glucose to UDP-glucuronate, an essential precursor for androgen inactivation by the prostate glucuronidation enzymes UGT2B15 and UGT2B17. UGDH expression is androgen stimulated, which increases the production of UDP-glucuronate and fuels UGT-catalyzed glucuronidation. In this study, we compared the glucuronidation potential and its impact on androgen-mediated gene expression in an isogenic LNCaP model for androgen-dependent versus castration-resistant prostate cancer. Despite significantly lower androgen-glucuronide output, LNCaP 81 castration-resistant tumor cells expressed higher levels of UGDH, UGT2B15, and UGT2B17. However, the magnitude of androgen-activated UGDH and prostate-specific antigen (PSA) expression, as well as the androgen receptor (AR)-dependent repression of UGT2B15 and UGT2B17, was blunted several-fold in these cells. Consistent with these results, the ligand-activated binding of AR to the PSA promoter and subsequent transcriptional activation were also significantly reduced in castration-resistant cells. Analysis of the UDP-sugar pools and flux through pathways downstream of UDP-glucuronate production revealed that these glucuronidation precursor metabolites were channeled through proteoglycan and glycosaminoglycan biosynthetic pathways, leading to increased surface expression of Notch1. Knockdown of UGDH diminished Notch1 and increased glucuronide output. Overall, these results support a model in which the aberrant partitioning of UDP-glucuronate and other UDP-sugars into alternative pathways during androgen deprivation contributes to the loss of prostate tumor cell androgen sensitivity by promoting altered cell surface proteoglycan expression.
前列腺上皮细胞部分通过失活和消除来控制雄激素的效力和可用性。UDP-葡萄糖脱氢酶(UGDH)催化UDP-葡萄糖依赖NAD(+)氧化为UDP-葡萄糖醛酸,这是前列腺葡萄糖醛酸化酶UGT2B15和UGT2B17使雄激素失活的必需前体。UGDH表达受雄激素刺激,这增加了UDP-葡萄糖醛酸的产生,并为UGT催化的葡萄糖醛酸化提供原料。在本研究中,我们在雄激素依赖与去势抵抗性前列腺癌的同基因LNCaP模型中比较了葡萄糖醛酸化潜力及其对雄激素介导的基因表达的影响。尽管雄激素-葡萄糖醛酸输出显著降低,但LNCaP 81去势抵抗性肿瘤细胞中UGDH、UGT2B15和UGT2B17的表达水平更高。然而,在这些细胞中,雄激素激活的UGDH和前列腺特异性抗原(PSA)表达的幅度,以及雄激素受体(AR)依赖性对UGT2B15和UGT2B17的抑制作用,均降低了数倍。与这些结果一致,在去势抵抗性细胞中,AR与PSA启动子的配体激活结合及随后的转录激活也显著降低。对UDP-糖库以及UDP-葡萄糖醛酸产生下游途径的通量分析表明,这些葡萄糖醛酸化前体代谢物通过蛋白聚糖和糖胺聚糖生物合成途径进行分流,导致Notch1的表面表达增加。敲低UGDH可减少Notch1并增加葡萄糖醛酸输出。总体而言,这些结果支持一种模型,即在雄激素剥夺期间,UDP-葡萄糖醛酸和其他UDP-糖异常分配到替代途径中,通过促进细胞表面蛋白聚糖表达改变,导致前列腺肿瘤细胞雄激素敏感性丧失。