P–284 Changes in protein expression due to metformin treatment and hyperinsulinemia in a human endometrial cancer cell line
P–284 Changes in protein expression due to metformin treatment and hyperinsulinemia in a human endometrial cancer cell line
复制标题
Pâ284 人子宫内膜癌细胞系中二甲双胍治疗和高胰岛素血症引起的蛋白质表达变化
DOI：
10.1093/humrep/deab130.283
复制
发表时间：
2021
影响因子：
6.1
通讯作者：
Germeyer
中科院分区：
文献类型：
--
作者：
Machado W Eber;Schmidt;Schroeder;Strowitzki;Germeyer
研究问题本研究的目的是通过蛋白质组学方法鉴定二甲双胍治疗对子宫内膜癌细胞影响的新靶蛋白/途径。概要答案研究了1,300种不同蛋白质的表达，其中80种蛋白质的变化最显著，并对其中一些进行了详细讨论。代谢性疾病如肥胖、II型糖尿病（T2 DM）和相关疾病（即多囊卵巢综合征（PCOS）、胰岛素抵抗）导致循环雌激素水平升高，从而促进EC的发生和进展。二甲双胍是一种胰岛素增敏双胍类药物，常用于治疗T2 DM，尤其是肥胖患者，在各种癌症类型（包括EC）中显示出抗癌作用。不同的蛋白质和途径被认为是潜在的靶点，但二甲双胍抗癌活性的潜在作用机制仍不完全清楚。研究设计、规模、持续时间在本体外研究中，EC细胞在5.5 mmol/L葡萄糖培养基中培养（补充10 nmol/L β-雌二醇（E2））和二甲双胍（0.5 mmol/L）、胰岛素（100 ng/mL）处理或不处理7 d。1,300种不同蛋白质的表达在细胞提取物中检测到的亲和蛋白质组学方法和治疗组之间的比较，以确定潜在的靶蛋白和途径，有助于抗癌作用的metformin.Participants/materials，setting，methodsThe研究进行了EC细胞系HEC-1A，代表绝经后模型与低E2敏感性。提取蛋白质，用BCA测定定量，并使用scioDiscover抗体微阵列分析蛋白质表达。样品之间蛋白质丰度的差异表示为log 2倍变化（log 2FC），对于显示出显著性的样品，|log 2光纤通道|≥ 0.5且调整后p ≤ 0.05。利用STRING和大卫数据库进行通路分析。主要结果和机会的作用数据显示，二甲双胍和胰岛素在本研究中靶向相似的通路，并且主要作用于与增殖、迁移和肿瘤免疫应答相关的蛋白。二甲双胍治疗或胰岛素补充可能以促肿瘤和抑肿瘤的方式影响这些途径。给出了80种受影响最大的蛋白质的结果，并详细讨论了几种蛋白质的检测到的表达变化对细胞的影响。所提供的数据有助于确定在EC中受二甲双胍治疗影响的潜在靶蛋白和途径，并允许更好地理解双胍类药物抗癌活性的作用机制。然而，需要进一步研究以证实二甲双胍给药后从所提供数据中得出的观察结果和结论，尤其是以有利方式调节的蛋白质，即AKT 3、CCND 2、CD 63、CD 81、GFAP、IL 5、IL 17 A、IRF 4、PI 3和VTCN 1。更多的蛋白质可能是感兴趣的，其中二甲双胍抵消了由高胰岛素血症诱导的不利影响。局限性，谨慎的原因该结果是从人类癌细胞系的体外研究中获得的，因此不能轻易地外推到患者。该发现的更广泛的含义：在高胰岛素血症环境的背景下，其他蛋白质可能是感兴趣的，即AMFR、CCND 2、CD 63、ERBB 3、EZR、GFAP、IRF 4.
Study questionThe aim of the study was to identify new target proteins/pathways that are affected by metformin treatment in endometrial cancer cells in a proteomic approach.Summary answerThe expression of 1,300 different proteins were investigated, of which 80 proteins with the most prominent changes were presented and some discussed in detail.What is known alreadyThe incidence of endometrial cancer (EC) has increased over the past years. Metabolic diseases such as obesity, type II diabetes mellitus (T2DM), and associated conditions (i.e. polycystic ovary syndrome (PCOS), insulin resistance) lead to elevated levels of circulating estrogens, which promote EC development and progression. Metformin, an insulin-sensitizing biguanide drug, commonly used in the treatment of T2DM, especially in obese patients, displayed anti-cancer effects in various cancer types, including EC. Different proteins and pathways have been suggested as potential targets, but the underlying mechanism of action of metformin’s anti-cancer activity is still not completely understood.Study design, size, durationIn the present in vitro study, EC cells were cultured in 5.5 mmol/L glucose medium (supplemented with 10 nmol/L ß-estradiol (E2)) and treated with metformin (0.5 mmol/L), insulin (100 ng/mL), or remained untreated for 7 d. The expression of 1,300 different proteins was detected in cellular extracts in an affinity proteomic approach and compared between the treatment groups in order to identify potential target proteins and pathways that contribute to the anti-cancer effects of metformin.Participants/materials, setting, methodsThe study was carried out with the EC cell line HEC–1A that represents a postmenopausal model with low E2 sensitivity. Proteins were extracted, quantified with the BCA assay, and protein expression was analyzed using the scioDiscover antibody microarray. Differences in protein abundance between samples were presented as log2-fold changes (log2FC) with significance for samples that displayed |log2FC| ≥ 0.5 and adjusted p ≤ 0.05. Pathway analysis was carried out with the STRING and DAVID databases.Main results and the role of chanceThe data revealed that metformin and insulin targeted similar pathways in the present study and mostly acted on proteins related to proliferation, migration and tumor immune response. These pathways may be affected in a tumor-promoting as well as a tumor-suppressing way by either metformin treatment or insulin supplementation. Results for the 80 most affected proteins were presented and the consequences for the cells resulting from the detected expression changes were discussed in detail for several proteins. The presented data helps identify potential target proteins and pathways affected by metformin treatment in EC and allows for a better understanding of the mechanism of action of the biguanide drug’s anti-cancer activity. However, further investigations are necessary to confirm the observations and conclusions drawn from the presented data after metformin administration, especially for proteins that were regulated in a favorable way, i.e. AKT3, CCND2, CD63, CD81, GFAP, IL5, IL17A, IRF4, PI3, and VTCN1. Further proteins might be of interest, where metformin counteracted unfavorable effects that have been induced by hyperinsulinemia.Limitations, reasons for cautionThe results were obtained from an in vitro study with human cancer cell lines, and thus cannot be easily extrapolated to patients.Wider implications of the findings: In the context of a hyperinsulinemic environment, further proteins might be of interest, i.e. AMFR, CCND2, CD63, ERBB3, EZR, GFAP, IRF4 …