Objective

To evaluate the effect of inhibition of histone deacetylases (HDACs) by suberoylanilide hydroxamic acid (SAHA) treatment of human uterine leiomyoma primary (HULP) cells in vitro on cell proliferation, cell cycle, extracellular matrix (ECM) formation, and transforming growth factor β3 (TGF-β3) signaling.

Design

Prospective study comparing uterine leiomyoma (UL) vs. adjacent myometrium (MM) tissue and cells with or without SAHA treatment.

Setting

Hospital and university laboratories.

Patient(s)

Women with UL without any hormone treatment.

Intervention(s)

Myomectomy or hysterectomy surgery in women for leiomyoma disease.

Main Outcome Measure(s)

HDAC activity was assessed by enzyme-linked immunosorbent assay, and gene expression was assessed by quantitative real-time polymerase chain reaction. Effects of SAHA on HULP cells were analyzed by CellTiter (Promega, Madison, Wisconsin), Western blot, and quantitative real-time polymerase chain reaction.

Result(s)

The expression of HDAC genes (HDAC1, fold change [FC] = 1.65; HDAC3, FC = 2.08; HDAC6, FC = 2.42) and activity (0.56 vs. 0.10 optical density [OD]/h/mg) was significantly increased in UL vs. MM tissue. SAHA decreased HDAC activity in HULP cells but not in MM cells. Cell viability significantly decreased in HULP cells (81.68% at 5 μM SAHA, 73.46% at 10 μM SAHA), but not in MM cells. Proliferating cell nuclear antigen expression was significantly inhibited in SAHA-treated HULP cells (5 μM SAHA, FC = 0.556; 10 μM SAHA, FC = 0.622). Cell cycle markers, including C-MYC (5 μM SAHA, FC = 0.828) and CCND1 (5 μM SAHA, FC = 0.583; 10 μM SAHA, FC = 0.482), were significantly down-regulated after SAHA treatment. SAHA significantly inhibited ECM protein expression, including FIBRONECTIN (5 μM SAHA, FC = 0.815; 10 μM SAHA, FC = 0.673) and COLLAGEN I (5 μM SAHA, FC = 0.599; 10 μM SAHA, FC = 0.635), in HULP cells. TGFβ3 and MMP9 gene expression was also significantly down-regulated by 10 μM SAHA (TGFβ3, FC = 0.596; MMP9, FC = 0.677).

Conclusion(s)

SAHA treatment inhibits cell proliferation, cell cycle, ECM formation, and TGF-β3 signaling in HULP cells, suggesting that histone deacetylation may be useful for treatment of UL.

中文翻译：

---

辛二酰苯胺异羟肟酸抑制组蛋白去乙酰化酶：治疗人子宫平滑肌瘤的治疗方法

客观的

评价辛二酰苯胺异羟肟酸 (SAHA) 处理体外人子宫平滑肌瘤 (HULP) 细胞抑制组蛋白去乙酰化酶 (HDAC) 对细胞增殖、细胞周期、细胞外基质 (ECM) 形成和转化生长因子 β3 的影响(TGF-β3) 信号传导。

设计

比较子宫平滑肌瘤 (UL) 与邻近子宫肌层 (MM) 组织和细胞的前瞻性研究，有无 SAHA 治疗。

环境

医院和大学实验室。

耐心）

未经任何激素治疗的 UL 女性。

干预措施

女性子宫肌瘤切除术或子宫切除术治疗平滑肌瘤疾病。

主要观察指标）

通过酶联免疫吸附测定评估HDAC活性，通过定量实时聚合酶链反应评估基因表达。通过 CellTiter (Promega, Madison, Wisconsin)、蛋白质印迹和定量实时聚合酶链反应分析 SAHA 对 HULP 细胞的影响。

结果）

HDAC 基因的表达（HDAC1，倍数变化 [FC] = 1.65；HDAC3，FC = 2.08；HDAC6，FC = 2.42）和活性（0.56 对 0.10 光密度 [OD]/h/mg）在 UL 中显着增加与 MM 组织相比。SAHA 降低了 HULP 细胞中的 HDAC 活性，但不降低 MM 细胞中的 HDAC 活性。HULP 细胞中的细胞活力显着降低（5 μM SAHA 时为 81.68%，10 μM SAHA 时为 73.46%），但 MM 细胞中没有。在 SAHA 处理的 HULP 细胞中增殖细胞核抗原表达受到显着抑制（5 μM SAHA，FC = 0.556；10 μM SAHA，FC = 0.622）。细胞周期标志物，包括C-MYC (5 μM SAHA, FC = 0.828) 和CCND1（5 μM SAHA，FC = 0.583；10 μM SAHA，FC = 0.482）在 SAHA 处理后显着下调。SAHA 在 HULP 细胞中显着抑制 ECM 蛋白表达，包括 FIBRONECTIN（5 μM SAHA，FC = 0.815；10 μM SAHA，FC = 0.673）和胶原蛋白 I（5 μM SAHA，FC = 0.599；10 μM SAHA，FC = 0.635） . TGFβ3和MMP9基因表达也被 10 μM SAHA 显着下调（TGFβ3，FC = 0.596；MMP9，FC = 0.677）。

结论

SAHA 治疗抑制 HULP 细胞中的细胞增殖、细胞周期、ECM 形成和 TGF-β3 信号传导，表明组蛋白去乙酰化可能对治疗 UL 有用。