The Lundquist Institute for Biomedical Innovation, Torrance, CA 90502, USA.
Department of Obstetrics and Gynecology, David Geffen School of Medicine at University of California, Los Angeles, CA 90095, USA.
Int J Mol Sci. 2024 Oct 27;25(21):11539. doi: 10.3390/ijms252111539.
Existing evidence indicates that LINCMD1 regulates muscle differentiation-related gene expression in skeletal muscle by acting as a miRNA sponge, though its role in leiomyoma development is still unknown. This study investigated LINCMD1's involvement in leiomyoma by analyzing paired myometrium and leiomyoma tissue samples (n = 34) from patients who had not received hormonal treatments for at least three months prior to surgery. Myometrium smooth muscle cells (MSMCs) were isolated, and gene expression of LINCMD1 and miR-135b was assessed via qRT-PCR, while luciferase assays determined the interaction between LINCMD1 and miR-135b. To examine the effects of LINCMD1 knockdown, siRNA transfection was applied to a 3D MSMC spheroid culture, followed by qRT-PCR and Western blot analyses of miR-135b, APC, β-Catenin and COL1A1 expression. The results showed that leiomyoma tissues had significantly reduced LINCMD1 mRNA levels, regardless of patient race or MED12 mutation status, while miR-135b levels were elevated compared to matched myometrium samples. Luciferase assays confirmed LINCMD1's role as a sponge for miR-135b. LINCMD1 knockdown in MSMC spheroids increased miR-135b levels, reduced APC expression, and led to β-Catenin accumulation and higher COL1A1 expression. These findings highlight LINCMD1 as a potential therapeutic target to modulate aberrant Wnt/β-Catenin signaling in leiomyoma.
现有证据表明,LINCMD1 通过作为 miRNA 海绵来调节骨骼肌中与肌肉分化相关的基因表达,尽管其在平滑肌瘤发展中的作用尚不清楚。本研究通过分析 34 例至少在手术前三个月未接受激素治疗的患者的配对子宫肌层和平滑肌瘤组织样本,研究了 LINCMD1 在平滑肌瘤中的作用。分离平滑肌细胞(MSMCs),并通过 qRT-PCR 评估 LINCMD1 和 miR-135b 的基因表达,而荧光素酶测定确定 LINCMD1 和 miR-135b 之间的相互作用。为了研究 LINCMD1 敲低的影响,将 siRNA 转染应用于 3D MSMC 球体培养物,然后进行 miR-135b、APC、β-Catenin 和 COL1A1 表达的 qRT-PCR 和 Western blot 分析。结果表明,无论患者种族或 MED12 突变状态如何,平滑肌瘤组织中的 LINCMD1 mRNA 水平均显著降低,而与匹配的子宫肌层样本相比,miR-135b 水平升高。荧光素酶测定证实了 LINCMD1 作为 miR-135b 海绵的作用。MSMC 球体中的 LINCMD1 敲低增加了 miR-135b 水平,降低了 APC 表达,导致 β-Catenin 积累和更高的 COL1A1 表达。这些发现强调了 LINCMD1 作为调节平滑肌瘤中异常 Wnt/β-Catenin 信号的潜在治疗靶标。