Simvastatin ameliorates altered mechanotransduction in uterine leiomyoma cells.
Simvastatin ameliorates altered mechanotransduction in uterine leiomyoma cells.
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辛伐他汀可以改善子宫平滑肌瘤细胞的机械转导改变。
DOI：
10.1016/j.ajog.2020.05.012
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发表时间：
2020-11
影响因子：
9.8
通讯作者：
Borahay MA
中科院分区：
文献类型：
--
作者：
Afrin S;Islam MS;Patzkowsky K;Malik M;Catherino WH;Segars JH;Borahay MA
子宫平滑肌瘤是女性生殖系统最常见的肿瘤，其特征是细胞外基质（ECM）的过度沉积和机械信号通路的根本改变。具体而言，这些改变影响了细胞外环境中对机械提示的正常动态响应状态。这些机械信号通过整合素、细胞膜受体转化为生化信号，包括细胞骨架信号传导途径，以维持机械稳态。平滑肌瘤细胞过度表达β1整合素和其他下游机械信号蛋白。我们曾报道过抗高血压药物辛伐他汀通过细胞、动物模型和流行病学研究具有抗平滑肌瘤作用。本研究的目的是检验辛伐他汀可能影响平滑肌瘤细胞中改变的机械传导的假设。这是一项基于实验室的实验研究。从约翰霍普金斯医院妇产科接受子宫切除术的五名患者中分离出原发性平滑肌瘤细胞。原代和永生化人平滑肌瘤（HuLM）细胞用辛伐他汀以递增浓度（0.001、0.01、0.1、1μM或对照）处理48 h。通过qRT-PCR、蛋白质印迹法和免疫荧光法定量参与机械信号传导的β1整联蛋白和ECM组分的蛋白质和mRNA水平。此外，我们使用下拉试验和凝胶收缩来检测辛伐他汀对RhoA活性的影响。我们发现，辛伐他汀显着降低了44%的β1整合素和60%的I型胶原蛋白的蛋白表达相比，未经处理的平滑肌瘤细胞。辛伐他汀处理的细胞减少FAK磷酸化下降到26-60%的控制，而它增加总FAK蛋白表达。使用RhoA下拉激活测定，我们观察到辛伐他汀处理的细胞与对照相比活性RhoA水平降低了45-85%。与受损的RhoA活化一致，辛伐他汀治疗减少了肿瘤凝胶收缩，其中凝胶面积比对照大122-153%。此外，辛伐他汀治疗导致参与β1整联蛋白下游信号传导的机械信号蛋白水平降低，包括AKAP 13、ROCK 1、MLCK和细胞周期蛋白D1。这项研究的结果表明，辛伐他汀可能的治疗作用，以恢复平滑肌瘤的机械信号转导的改变状态。总的来说，这些发现与以前的流行病学和其他报告一致，并支持临床试验的必要性。
Uterine leiomyomas, the most common tumors of the female reproductive system, feature excessive deposition of disordered stiff extracellular matrix (ECM) and fundamentally altered mechanical signaling pathways. Specifically, these alterations impact the normal dynamic state of responsiveness to mechanical cues in the extracellular environment. These mechanical cues are converted via integrins, cell membrane receptors, to biochemical signals including cytoskeletal signaling pathways to maintain mechanical homeostasis. Leiomyoma cells overexpress β1 integrin and other downstream mechanical signaling proteins. We previously reported that simvastatin, an anti-hyperlipidemic drug, has anti-leiomyoma effects through cellular, animal model and epidemiologic studies. The purpose of this study was to examine the hypothesis that simvastatin might influence altered mechanotransduction in leiomyoma cells. This is a laboratory-based experimental study. Primary leiomyoma cells were isolated from five patients who underwent hysterectomy at the Department of Gynecology and Obstetrics at the Johns Hopkins Hospital. Primary and immortalized human leiomyoma (HuLM) cells were treated with simvastatin at increasing concentrations (0.001, 0.01, 0.1, 1μM or control) for 48 h. Protein and mRNA levels of β1 integrin and ECM components involved in mechanical signaling were quantified by qRT-PCR, western blotting, and immunofluorescence. In addition, we examined the effect of simvastatin on RhoA activity using pull-down assay and gel contraction. We found that simvastatin significantly reduced the protein expression of β1 integrin by 44% and type I collagen by 60% compared to untreated leiomyoma cells. Simvastatin-treated cells reduced phosphorylation of FAK down to 26-60% of control whereas it increased total FAK protein expression. Using a RhoA pull-down activation assay, we observed that simvastatin-treated cells had reduced levels of active RhoA by 45-85% compared to control. Consistent with impaired RhoA activation, simvastatin treatment reduced tumor gel contraction where gel area was 122-153% larger compared to control. Furthermore, simvastatin treatment led to reduced levels of mechanical signaling proteins involved in β1 integrin downstream signaling, including AKAP13, ROCK1, MLCK, and cyclin D1. The results of this study suggest a possible therapeutic role for simvastatin to restore the altered state of mechanotransduction signaling in leiomyoma. Collectively, the findings are aligned with previous epidemiologic and other reports and support the need for clinical trials.
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