Acetyl-L-Carnitine downregulates invasion (CXCR4/CXCL12, MMP-9) and angiogenesis (VEGF, CXCL8) pathways in prostate cancer cells: rationale for prevention and interception strategies
Acetyl-L-Carnitine downregulates invasion (CXCR4/CXCL12, MMP-9) and angiogenesis (VEGF, CXCL8) pathways in prostate cancer cells: rationale for prevention and interception strategies
复制标题
DOI：
10.1186/s13046-019-1461-z
复制
发表时间：
2019-11-12
影响因子：
11.3
通讯作者：
Noonan, Douglas M.
中科院分区：
文献类型：
--
作者：
Baci, Denisa;Bruno, Antonino;Noonan, Douglas M.
背景前列腺癌（PCa）是全世界男性癌症相关死亡的主要原因。炎症加剧和血管生成已在很大程度上被证明有助于PCa进展。各种天然存在的化合物和膳食补充剂具有抗氧化、抗炎和抗血管生成活性，代表了在化学预防环境中靶向控制PCa进展和血管生成的异常细胞因子/趋化因子产生的有效化合物。使用质谱分析前列腺癌患者的血清样本，我们以前发现在非癌症个体中肉毒碱的水平较高，这表明了保护作用。在这里，我们研究了乙酰-L-肉碱（ALCAR）在体外和体内干扰前列腺癌进展和血管生成的关键功能特性的能力，并确定了ALCAR调节的靶分子。方法体外研究ALCAR对4种不同的前列腺癌细胞系（PC-3、DU-145、LNCaP、22 Rv 1）和良性前列腺增生（BPH）细胞系的化学预防/血管预防活性。流式细胞术（FC）检测ALCAR诱导细胞凋亡和细胞周期阻滞的作用。进行细胞粘附、迁移和侵袭（Boyden小室）的功能分析。通过FC评估ALCAR对表面抗原受体（趋化因子）和细胞内细胞因子产生的调节。通过多重免疫测定检测促血管生成因子的释放。使用肿瘤异种移植物研究了ALCAR对体内PCa细胞生长的影响。结果ALCAR可抑制前列腺癌细胞增殖，诱导凋亡，抑制促炎细胞因子（TNF-α和IFN-γ）和趋化因子CCL 2、CXCL 12及趋化轴相关受体CXCR 4的产生，降低前列腺癌细胞和前列腺增生细胞的粘附、迁移和侵袭能力。ALCAR通过减少促血管生成因子（VEGF、CXCL 8、CCL 2、血管生成素）和金属蛋白酶MMP-9的产生/释放，对PCa发挥血管预防活性。与未处理细胞相比，将内皮细胞暴露于来自用ALCAR预处理的PCa细胞的条件培养基抑制了CXCR 4、CXCR 1、CXCR 2和CCR 2的表达。对异种移植有两种不同PCa细胞系的小鼠口服（饮用水）ALCAR导致体内肿瘤细胞生长减少。结论我们的研究结果强调了ALCAR通过减少几种关键趋化因子、细胞因子和MMP 9的产生来下调前列腺癌细胞的生长、粘附、迁移和侵袭的能力。ALCAR是一种广泛分布的膳食补充剂，我们的研究结果为研究ALCAR作为前列腺癌高危受试者化学预防方法的可能分子提供了依据。我们建议ALCAR作为一种新的可能的“重新利用的药物”，用于癌症预防和拦截，类似于阿司匹林，二甲双胍或β-受体阻滞剂。
Background Prostate cancer (PCa) is a leading cause of cancer-related death in males worldwide. Exacerbated inflammation and angiogenesis have been largely demonstrated to contribute to PCa progression. Diverse naturally occurring compounds and dietary supplements are endowed with anti-oxidant, anti-inflammatory and anti-angiogenic activities, representing valid compounds to target the aberrant cytokine/chemokine production governing PCa progression and angiogenesis, in a chemopreventive setting. Using mass spectrometry analysis on serum samples of prostate cancer patients, we have previously found higher levels of carnitines in non-cancer individuals, suggesting a protective role. Here we investigated the ability of Acetyl-L-carnitine (ALCAR) to interfere with key functional properties of prostate cancer progression and angiogenesis in vitro and in vivo and identified target molecules modulated by ALCAR. Methods The chemopreventive/angiopreventive activities ALCAR were investigated in vitro on four different prostate cancer (PCa) cell lines (PC-3, DU-145, LNCaP, 22Rv1) and a benign prostatic hyperplasia (BPH) cell line. The effects of ALCAR on the induction of apoptosis and cell cycle arrest were investigated by flow cytometry (FC). Functional analysis of cell adhesion, migration and invasion (Boyden chambers) were performed. ALCAR modulation of surface antigen receptor (chemokines) and intracellular cytokine production was assessed by FC. The release of pro-angiogenic factors was detected by a multiplex immunoassay. The effects of ALCAR on PCa cell growth in vivo was investigated using tumour xenografts. Results We found that ALCAR reduces cell proliferation, induces apoptosis, hinders the production of pro inflammatory cytokines (TNF-alpha and IFN-gamma) and of chemokines CCL2, CXCL12 and receptor CXCR4 involved in the chemotactic axis and impairs the adhesion, migration and invasion capabilities of PCa and BPH cells in vitro. ALCAR exerts angiopreventive activities on PCa by reducing production/release of pro angiogenic factors (VEGF, CXCL8, CCL2, angiogenin) and metalloprotease MMP-9. Exposure of endothelial cells to conditioned media from PCa cells, pre-treated with ALCAR, inhibited the expression of CXCR4, CXCR1, CXCR2 and CCR2 compared to those from untreated cells. Oral administration (drinking water) of ALCAR to mice xenografted with two different PCa cell lines, resulted in reduced tumour cell growth in vivo. Conclusions Our results highlight the capability of ALCAR to down-modulate growth, adhesion, migration and invasion of prostate cancer cells, by reducing the production of several crucial chemokines, cytokines and MMP9. ALCAR is a widely diffused dietary supplements and our findings provide a rational for studying ALCAR as a possible molecule for chemoprevention approaches in subjects at high risk to develop prostate cancer. We propose ALCAR as a new possible "repurposed agent' for cancer prevention and interception, similar to aspirin, metformin or beta-blockers.