Uniformed Services University of the Health Sciences, Bethesda, Maryland; Program in Reproductive Endocrinology and Gynecology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland.
Uniformed Services University of the Health Sciences, Bethesda, Maryland.
Fertil Steril. 2020 Jan;113(1):176-186. doi: 10.1016/j.fertnstert.2019.08.090.
To characterize the role of steroid hormone and antihormone exposure on neurotrimin (NTM) expression in human leiomyoma and myometrial tissue and cells.
Laboratory study of placebo and ulipristal acetate (UPA)-treated patient tissue. In vitro assessment of immortalized myometrial and leiomyoma cell lines after hormone and antihormone exposure.
Academic research center.
PATIENT(S): Not applicable.
INTERVENTIONS(S): Exposure of leiomyoma cell lines to 17β-E, medroxyprogesterone acetate (MPA), UPA, and fulvestrant.
MAIN OUTCOME MEASURE(S): Messenger RNA expression quantified with the use of RNASeq analysis and quantitative real-time polymerase chain reaction (qRT-PCR). Protein levels quantified by means of Western blot analysis. Immunohistochemistry (IHC) on placebo- and UPA-treated patient uterine tissue specimens.
RESULT(S): Expression of NTM in human uterine leiomyoma specimens according to RNASeq was increased compared with myometrium (5.22 ± 0.57-fold), which was confirmed with the use of qRT-PCR (1.95 ± 0.05). Furthermore, NTM protein was elevated in leiomyoma tissue compared with matched myometrium (2.799 ± 0.575). IHC revealed increased staining intensity in leiomyoma surgical specimens compared with matched myometrium of placebo patients. Western blot analysis in immortalized leiomyoma cell lines demonstrated an up-regulation of NTM protein expression (2.4 ± 0.04). Treatment of leiomyoma cell lines with 17β-E yielded a 1.98 ± 0.11-fold increase in NTM protein expression; however, treatment with fulvestrant showed no significant change compared with control. Leiomyoma cell lines demonstrated a 1.91 ± 0.97-fold increase in NTM protein expression after progesterone treatment. RNASeq analysis demonstrated a reduced expression in patient leiomyoma after UPA treatment (0.75 ± 0.14). Treatment of leiomyoma cells with UPA demonstrated a reduced total NTM protein amount (0.54 ± 0.31) in patients, which was confirmed with the use of IHC (UPA10 147.2 ± 9.40, UPA20 182.8 ± 8.98). In vitro studies with UPA treatment revealed a concentration-dependent effect that supported these findings.
CONCLUSION(S): NTM, a neural cell adhesion molecule, is increased in leiomyoma compared with myometrium in patient tissue and in vitro models after estrogen and progesterone treatment. Down-regulation of expression occurs after UPA treatment, but not after fulvestrant exposure.
NCT00290251.
描述甾体激素和抗激素暴露对人子宫肌瘤和子宫肌组织和细胞中神经生长调节蛋白（NTM）表达的作用。
安慰剂和屈螺酮（UPA）治疗患者组织的实验室研究。激素和抗激素暴露后对永生化子宫肌和子宫肌瘤细胞系的体外评估。
学术研究中心。
不适用。
子宫肌瘤细胞系暴露于 17β-E、甲羟孕酮（MPA）、UPA 和氟维司群。
使用 RNAseq 分析和实时定量聚合酶链反应（qRT-PCR）定量信使 RNA 表达。Western blot 分析定量蛋白质水平。安慰剂和 UPA 治疗的患者子宫组织标本的免疫组织化学（IHC）。
根据 RNAseq，与子宫肌相比，人子宫肌瘤标本中 NTM 的表达增加（5.22 ± 0.57 倍），这通过 qRT-PCR 得到了证实（1.95 ± 0.05）。此外，与匹配的子宫肌相比，子宫肌瘤组织中 NTM 蛋白升高（2.799 ± 0.575）。IHC 显示与安慰剂患者匹配的子宫肌相比，子宫肌瘤手术标本中染色强度增加。Western blot 分析在永生化子宫肌瘤细胞系中显示 NTM 蛋白表达上调（2.4 ± 0.04）。17β-E 处理子宫肌瘤细胞系导致 NTM 蛋白表达增加 1.98 ± 0.11 倍；然而，与对照相比，用氟维司群处理没有显示出显著变化。孕激素处理后，子宫肌瘤细胞系中 NTM 蛋白表达增加 1.91 ± 0.97 倍。RNAseq 分析显示 UPA 治疗后患者子宫肌瘤表达减少（0.75 ± 0.14）。UPA 处理子宫肌瘤细胞显示患者 NTM 总蛋白量减少（0.54 ± 0.31），这通过 IHC 得到证实（UPA10 147.2 ± 9.40，UPA20 182.8 ± 8.98）。UPA 治疗的体外研究显示出浓度依赖性效应，支持了这些发现。
NTM，一种神经细胞粘附分子，在患者组织和雌激素和孕激素处理后的体外模型中，与子宫肌相比，在子宫肌瘤中增加。UPA 治疗后表达下调，但氟维司群暴露后没有下调。
NCT00290251。
