尿路上皮癌中组蛋白去乙酰化酶（HDAC）表达模式的改变和 HDAC 抑制剂的活性。

Changes in histone deacetylase (HDAC) expression patterns and activity of HDAC inhibitors in urothelial cancers.

机构信息

Department of Urology, Heinrich-Heine-University, Medical Faculty, Düsseldorf, Germany.

出版信息

Urol Oncol. 2013 Nov;31(8):1770-9. doi: 10.1016/j.urolonc.2012.06.015. Epub 2012 Sep 1.

DOI:10.1016/j.urolonc.2012.06.015

PMID:22944197

Abstract

OBJECTIVE

To determine histone deacetylase (HDAC) isoenzyme expression patterns in urothelial cancer tissues and cell lines and investigate their potential to predict the efficacy of the HDAC inhibitor vorinostat.

MATERIALS AND METHODS

Expression of HDAC mRNAs was determined by quantitative RT-PCR in 18 urothelial cancer cell lines (UCC), normal uroepithelial controls (NUC), 24 urothelial cancer tissues, and 12 benign controls. Results were compared with published microarray data. Effects of pan-HDAC inhibitor vorinostat and on UCCs were determined by viability and apoptosis assays, cell cycle analysis, and measurements of p21(CIP1), thymidylate synthase (TS), and EZH2. In addition, protein expression levels of HDACs were investigated in UCCs.

RESULTS

Prominent changes in UCCs were HDAC2 and/or HDAC8 up-regulation in 11 of 18 cell lines and decreased expression of HDAC4, HDAC5, and/or HDAC7 mRNA in 15 of 18 cell lines. In cancer tissues, HDAC8 was likewise significantly up-regulated (P = 0.002), whereas HDAC2 up-regulation was detected only in a subset of tumors (9/24, P = 0.085). Overexpression of HDAC2 and HDAC8 mRNA did not correspond with the protein level. Vorinostat induced G2/M arrest, an increase in the sub-G1 fraction, up-regulation of p21, and down-regulation of TS in all UCC. Effects on EZH2 and PARP cleavage as well as activation of caspase 3/7 differed between cell lines. Associations between the overall sensitivity to the pan-HDACi vorinostat and overexpression of HDAC2 and HDAC8 mRNA were not observed.

CONCLUSIONS

In urothelial cancer, up-regulation of HDAC2 and HDAC8 and down-regulation of HDAC4, HDAC5, and HDAC7 mRNA are common findings. The treatment effect of the pan-HDAC inhibitor vorinostat was variable in UCCs and up-regulation of HDAC2 and HDAC8 was not predictive for treatment response. Whether selective targeting of HDAC2, HDAC8, or other HDACs deregulated in urothelial cancer (e.g., HDAC4, HDAC5, and HDAC7) result in a more consistent treatment response needs further investigation.

摘要

目的

确定尿路上皮癌组织和细胞系中组蛋白去乙酰化酶（HDAC）同工酶的表达模式，并研究其预测 HDAC 抑制剂伏立诺他疗效的潜力。

材料与方法

采用定量 RT-PCR 法检测 18 株尿路上皮癌细胞系（UCC）、正常尿上皮对照（NUC）、24 份尿路上皮癌组织和 12 份良性对照中 HDACmRNA 的表达。结果与已发表的微阵列数据进行比较。采用细胞活力和凋亡测定、细胞周期分析以及检测 p21（CIP1）、胸苷酸合成酶（TS）和 EZH2，研究泛 HDAC 抑制剂伏立诺他对 UCC 的作用。此外，还研究了 UCC 中 HDAC 的蛋白表达水平。

结果

在 18 株细胞系中，有 11 株细胞系的 HDAC2 和/或 HDAC8 上调，有 15 株细胞系的 HDAC4、HDAC5 和/或 HDAC7mRNA 表达下调；在癌组织中，HDAC8 也明显上调（P = 0.002），而 HDAC2 上调仅在部分肿瘤中检测到（9/24，P = 0.085）。HDAC2 和 HDAC8mRNA 的过表达与蛋白水平不对应。伏立诺他诱导所有 UCC 的 G2/M 期阻滞、亚 G1 区增加、p21 上调和 TS 下调。细胞系之间在 EZH2 和 PARP 切割以及 caspase 3/7 激活的作用存在差异。泛 HDACi 伏立诺他的总体敏感性与 HDAC2 和 HDAC8mRNA 的过表达之间无关联。

结论

在尿路上皮癌中，HDAC2 和 HDAC8 的上调以及 HDAC4、HDAC5 和 HDAC7mRNA 的下调是常见的发现。泛 HDAC 抑制剂伏立诺他对 UCC 的治疗效果各不相同，HDAC2 和 HDAC8 的上调不能预测治疗反应。在尿路上皮癌中，靶向调控 HDAC2、HDAC8 或其他失调的 HDAC（如 HDAC4、HDAC5 和 HDAC7）是否会导致更一致的治疗反应，需要进一步研究。