The effect of the readthrough acetylcholinesterase variant (AChE-R) on uterine muscle and leiomyomas
The effect of the readthrough acetylcholinesterase variant (AChE-R) on uterine muscle and leiomyomas
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DOI：
10.1093/molehr/gam010
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发表时间：
2007-05-01
影响因子：
4
通讯作者：
Deutsch, Varda
中科院分区：
文献类型：
--
作者：
Grisaru, Dan;Keidar, Ran;Deutsch, Varda
乙酰胆碱信号和乙酰胆碱酯酶（AChE）功能是肌肉发育的关键因素。在体内和体外评估了刺激依赖性读透AChE变体AChE- r对平滑肌瘤和正常肌层增殖的影响。组织学准备和细胞培养是在子宫切除术或子宫肌瘤切除术中获得的，包括平滑肌瘤样本和相邻的正常子宫肌肉作为对照。使用与人AChE- r转录物互补的AChE cRNA探针进行原位杂交。抗AChE-R变异的抗体用于免疫组织化学染色。为了确定AChE-R对子宫肌细胞培养的生物学功能，我们使用了一种合成肽，代表了AChE-R （ARP）的潜在可切割的形态活性c端。通过掺入5'-溴-2-脱氧尿苷（BrDU）评估细胞增殖。与正常肌层相比，平滑肌瘤表达了过量的AChE-R mRNA和AChE-R蛋白。来源于平滑肌瘤的细胞培养比来源于相邻肌层的细胞培养增殖明显快（在BrDU掺入时P = 0.027）。添加ARP （2-200 nM）引起平滑肌瘤和肌层细胞培养物增殖的剂量依赖性降低。对肌层的影响达到了统计学意义（在20和200 nM， P = 0.02），而快速增殖的原代培养物的变异性很高，在平滑肌瘤培养物中不具有统计学意义。AChE-R参与肌层的增殖。ARP对肌层的抑制作用可能提示其未来的治疗作用。
Acetylcholine signaling and acetyleholinesterase (AChE) function(s) are pivotal elements in muscle development. The effects of the stimulus-dependent readthrough AChE variant, AChE-R, on leiomyomas and normal myometrium proliferation were assessed in vivo and in vitro. Histological preparations and cell cultures therefrom were obtained during hysterectomies or myomectomies and included both the leiomyoma sample and the adjacent normal uterine muscle as control. In situ hybridization procedures were performed using AChE cRNA probes complementary to the human AChE-R transcript. Antibodies against the AChE-R variant served for immun ohistochemical staining. To determine the biological function of AChE-R on the uterine muscle cell cultures, we used a synthetic peptide representing the potentially cleavable morphogenically active C-terminus of AChE-R (ARP). Cell proliferation was assessed using the incorporation of 5'-bromo-2-deoxyuridine (BrDU). Leiomyomas expressed an excess of AChE-R mRNA and the AChE-R protein compared with the normal myometrium. Cell cultures originating from leiomyomas proliferated significantly faster than cultures from the adjacent myometrium (P = 0.027 at BrDU incorporation). Addition of ARP (2-200 nM) caused a dose-dependent decrease in the proliferation of cell cultures from both leiomyomas and the myometrium. The effect on the myometrium reached statistical significance (at 20 and 200 nM, P = 0.02), whereas the variability of the rapidly proliferating primary cultures was high and precluded statistical significance in the leiomyoma cultures. AChE-R is involved in the proliferation of the myometrium. The inhibitory effect of ARP on the myometrium may suggest a future therapeutic role of ARP.