MT1-MMP 在血管内膜增生过程中雌激素介导的细胞活动中的作用。

Role of MT1-MMP in estrogen-mediated cellular processes of intimal hyperplasia.

机构信息

Department of Surgery, University of Tennessee Graduate School of Medicine, Knoxville, Tennessee 37920, USA.

出版信息

J Surg Res. 2012 Apr;173(2):224-31. doi: 10.1016/j.jss.2011.05.037. Epub 2011 Jun 23.

DOI:10.1016/j.jss.2011.05.037

PMID:21777927

Abstract

BACKGROUND

Hormone replacement therapy increases intimal hyperplasia (IH) following vascular intervention. Matrix metalloproteinases (MMPs) play a role in IH development. We have shown estrogen up-regulates MT1-MMP expression, a transmembrane protein that activates MMP-2, and increases vascular smooth muscle cell (VSMC) collagen invasion via increased MMP-2 activity. Here we hypothesize inhibition of MT1-MMP will prevent hormonally-stimulated increased MMP-2 activation and the downstream cellular processes of IH pathogenesis.

METHODS

VSMCs from a postmenopausal donor were transfected with MT1-MMP or negative control siRNAs, treated with estrogen (Est), analyzed by q-PCR, Western blot, zymography, migration, invasion, and proliferation assays.

RESULTS

Est treatment of MT1-MMP silenced cells still resulted in increased MT1-MMP expression (C = 41% ± 4%; Est = 52% ± 2%; P < 0.05). Silencing of MT1-MMP decreased basal MMP-2 activity (nonsilenced = 100%; MT1-silenced = 87% ± 3%; P < 0.05) but had no effect on basal invasion or proliferation. Est treatment of MT1-MMP silenced cells still resulted in increased MMP-2 activity (C = 87% ± 3%; Est = 101% ± 4%; P < 0.05) and invasion (C = 89% ± 6%; Est = 109% ± 3%; P < 0.05) compared with MT1-MMP silenced control cells. However, silencing of MT1-MMP did inhibit Est- and serum-stimulated proliferation (C = 106% ± 18%; Est = 104% ± 16%; FBS = 121% ± 24%; P = NS).

CONCLUSION

Silencing of MT1-MMP in aged VSMCs results in impaired but not complete inhibition of basal and Est-stimulated increases in MMP-2 activity. Other mechanisms appear to be playing a role in hormonally-regulated cellular processes of IH pathogenesis. Future studies will target other signaling cascades, with the goal of identifying mechanisms responsible for hormonally-modulated unbalanced MMPs. In vivo manipulation of the expression patterns of MT1-MMP will be examined for the prevention of IH in animal models of vascular disease.

摘要

背景

激素替代疗法会增加血管介入后的内膜增生（IH）。基质金属蛋白酶（MMPs）在 IH 发展中起作用。我们已经表明，雌激素上调 MT1-MMP 的表达，MT1-MMP 是一种激活 MMP-2 的跨膜蛋白，并通过增加 MMP-2 活性增加血管平滑肌细胞（VSMC）胶原浸润。在这里，我们假设抑制 MT1-MMP 将防止激素刺激的 MMP-2 激活和 IH 发病机制的下游细胞过程。

方法

用 MT1-MMP 或阴性对照 siRNA 转染绝经后供体的 VSMCs，用雌激素（Est）处理，通过 q-PCR、Western blot、酶谱、迁移、侵袭和增殖试验进行分析。

结果

尽管 MT1-MMP 沉默细胞仍接受雌激素处理，但 MT1-MMP 的表达仍增加（C = 41% ± 4%；Est = 52% ± 2%；P < 0.05）。MT1-MMP 沉默降低了基础 MMP-2 活性（非沉默 = 100%；MT1 沉默 = 87% ± 3%；P < 0.05），但对基础侵袭或增殖没有影响。MT1-MMP 沉默细胞的雌激素处理仍导致 MMP-2 活性增加（C = 87% ± 3%；Est = 101% ± 4%；P < 0.05）和侵袭（C = 89% ± 6%；Est = 109% ± 3%；P < 0.05），与 MT1-MMP 沉默对照细胞相比。然而，MT1-MMP 的沉默确实抑制了 Est 和血清刺激的增殖（C = 106% ± 18%；Est = 104% ± 16%；FBS = 121% ± 24%；P = NS）。

结论

在衰老的 VSMCs 中沉默 MT1-MMP 导致基础和 Est 刺激的 MMP-2 活性增加受损，但不完全抑制。其他机制似乎在 IH 发病机制的激素调节细胞过程中起作用。未来的研究将针对其他信号级联，目标是确定负责激素调节的 MMP 失衡的机制。将研究体内对 MT1-MMP 表达模式的操作，以防止血管疾病动物模型中的 IH。