Histone deacetylase inhibition by suberoylanilide hydroxamic acid: a therapeutic approach to treat human uterine leiomyoma
Histone deacetylase inhibition by suberoylanilide hydroxamic acid: a therapeutic approach to treat human uterine leiomyoma
复制标题
DOI：
10.1016/j.fertnstert.2021.10.012
复制
发表时间：
2022-02-03
影响因子：
6.7
通讯作者：
Ferrero, Hortensia
中科院分区：
文献类型：
--
作者：
Carbajo-Garcia, Maria Cristina;Garcia-Alcazar, Zaira;Ferrero, Hortensia
目的：评价辛二酰苯胺异羟肟酸（SAHA）体外处理人子宫平滑肌瘤原代（HULP）细胞抑制组蛋白脱乙酰酶（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/53（TGF-β 53）信号传导的影响。比较子宫平滑肌瘤（UL）与相邻子宫肌层（MM）组织和细胞（有或无SAHA治疗）的前瞻性研究。设置：医院和大学实验室。患者：未接受任何激素治疗的UL妇女。干预措施：子宫肌瘤切除术或子宫切除术治疗子宫肌瘤疾病的妇女。主要结果测量：HDAC活性通过酶联免疫吸附测定进行评估，基因表达通过定量实时聚合酶链反应进行评估。CellTiter分析SAHA对HULP细胞的作用（Promega，麦迪逊，威斯康星州）、蛋白质印迹和定量实时聚合酶链反应。（HDAC 1，倍数变化[FC] = 1.65; HDAC 3，FC = 2.08; HDAC 6，FC = 2.42）和活性（0.56对0.10光密度[OD]/h/mg）在UL对MM组织中显著增加。SAHA降低HULP细胞中的HDAC活性，但不降低MM细胞中的HDAC活性。HULP细胞中细胞活力显著降低（5 μ M SAHA时为81.68%，10 μ M 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被显著抑制（5 μ M SAHA，FC = 0.556; 10 μ M SAHA，FC = 0.622）。细胞周期标志物，包括C-MYC（5 μ M SAHA，FC = 0.828）和CCND 1（5 μ M SAHA，FC = 0.583; 10 μ M SAHA，FC = 0.482），在SAHA处理后显著下调。在HULP细胞中，SAHA显著抑制ECM蛋白表达，包括纤维连接蛋白（5 μ M SAHA，FC = 0.815; 10 μ M SAHA，FC = 0.673）和胶原I（5 μ M SAHA，FC = 0.599; 10 μ M SAHA，FC = 0.635）。10 μ M SAHA可显著下调HULP细胞TGF-53和MMP-9基因表达（TGF-53，FC = 0.596; MMP-9，FC = 0.677）。结论：SAHA可抑制HULP细胞增殖、细胞周期、ECM形成和TGF-53信号通路，提示组蛋白去乙酰化可能对UL的治疗有一定作用。（Fertil Sterile 2022;117：433-43. (c)2021年由美国生殖医学协会。
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 /53 (TGF-/53) 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 mu M SAHA, 73.46% at 10 mu M SAHA), but not in MM cells. Proliferating cell nuclear antigen expression was significantly inhibited in SAHA-treated HULP cells (5 mu M SAHA, FC = 0.556; 10 mu M SAHA, FC = 0.622). Cell cycle markers, including C-MYC (5 mu M SAHA, FC = 0.828) and CCND1 (5 mu M SAHA, FC = 0.583; 10 mu M SAHA, FC = 0.482), were significantly down-regulated after SAHA treatment. SAHA significantly inhibited ECM protein expression, including FIBRONECTIN (5 mu M SAHA, FC = 0.815; 10 mu M SAHA, FC = 0.673) and COLLAGEN I (5 mu M SAHA, FC = 0.599; 10 mu M SAHA, FC = 0.635), in HULP cells. TGF/53 and MMP9 gene expression was also significantly down-regulated by 10 mu M SAHA (TGF/53, FC = 0.596; MMP9, FC = 0.677).Conclusion(s): SAHA treatment inhibits cell proliferation, cell cycle, ECM formation, and TGF-/53 signaling in HULP cells, suggesting that histone deacetylation may be useful for treatment of UL. (Fertil Sterile 2022;117:433-43. (c) 2021 by American Society for Reproductive Medicine.)