Calebin-A 通过激活组蛋白乙酰转移酶诱导恶性外周神经鞘瘤细胞死亡。
Calebin-A induced death of malignant peripheral nerve sheath tumor cells by activation of histone acetyltransferase.
机构信息
Department of Neurology, National Taiwan University Hospital, Taipei 10012, Taiwan; Department of Medical Genetics, National Taiwan University Hospital, Taipei 10012, Taiwan.
Department of Neurology, National Taiwan University Hospital, Taipei 10012, Taiwan.
出版信息
Phytomedicine. 2019 Apr;57:377-384. doi: 10.1016/j.phymed.2019.01.001. Epub 2019 Jan 3.
BACKGROUND
Neurofibromatosis type 1 (NF1) is one of the most common hereditary neurocutaneous disorders. The malignant peripheral nerve sheath tumor (MPNST), transformed from NF1 related plexiform neurofibroma, is a rapidly growing and highly invasive tumor. No effective chemotherapeutic agent is currently available. Calebin-A is a derivative from turmeric Curcuma longa. Given the anti-inflammatory and anticancer potentials of curcumin, whether Calebin-A also had the tumoricidal effect upon MPNST cells is still elusive.
PURPOSE
To determine whether Calebin-A has the potential for anti-MPNST effect.
METHODS
The MTT and FACS analysis of normal Schwann (HSC) and MPNST cells have been employed to determine the tumoricidal effect of Calebin-A. The expression of the signal pathway molecules was assessed by Western blotting. The CHIP with quantitative PCR assay was performed to quantify the promoter DNA binding to acetylated histone 3 (acetyl H3). The enzyme activities of histone acetyltransferase (HAT) and deacetylase (HDAC) have been evaluated by commercial kits. The measurements of tumor size of the xenograft mouse model were also performed.
RESULTS
Calebin-A inhibited the proliferation of MPNST and primary neurofibroma cells in a dose-dependent manner. The flow cytometry analysis of the MPNST cells after treatment of 25 μm of Calebin-A demonstrated an increase of population in the G0/G1 phase but decrease in G2/M phase. Before treatment, the expression of Axl, Tyro3, and acetyl H3 was significantly higher in MPNST cells when compared to HSC. The expression of phosphorylated-AKT, -ERK1/2, survivin, hTERT, and acetyl H3 proteins were reduced after treatment. The CHIP assay shows the promoter DNA copies of survivin (BRIC5) and hTERT genes are significantly reduced post-treatment. The enzyme activity of HAT was significantly reduced, but not that of HDAC. Two HAT inhibitors, epigallocatechin-3-gallate (EGCG) and anacardic acid (AA) have also demonstrated a significant inhibitory effect on MPNST cells. Finally, the measurements of tumor size showed a significant reduction of the xenograft tumors after treatment of Calebin-A.
CONCLUSION
Both in vitro and in vivo studies showed Calebin-A could inhibit the proliferation of MPNST with suppression of survivin and hTERT. The reduced expression of these two factors might be through the epigenetic histone modification resulting from the decreased activity of HAT.
背景
神经纤维瘤病 1 型（NF1）是最常见的遗传性神经皮肤疾病之一。恶性外周神经鞘瘤（MPNST）是由 NF1 相关丛状神经纤维瘤转化而来的一种快速生长且高度侵袭性的肿瘤。目前尚无有效的化疗药物。姜黄素是姜黄的一种衍生物。鉴于姜黄素具有抗炎和抗癌作用，姜黄素衍生物 Calebin-A 是否对 MPNST 细胞也具有杀伤作用仍不清楚。
目的
确定 Calebin-A 是否具有抗 MPNST 作用。
方法
采用 MTT 和 FACS 分析法检测 Calebin-A 对正常许旺（HSC）细胞和 MPNST 细胞的杀伤作用。采用 Western blot 法检测信号通路分子的表达。采用定量 PCR 芯片检测乙酰化组蛋白 3（acetyl H3）与启动子 DNA 的结合。采用商业试剂盒评估组蛋白乙酰转移酶（HAT）和去乙酰化酶（HDAC）的酶活性。还对异种移植小鼠模型的肿瘤大小进行了测量。
结果
Calebin-A 呈剂量依赖性抑制 MPNST 和原发性神经纤维瘤细胞的增殖。用 25 µm Calebin-A 处理 MPNST 细胞后的流式细胞术分析显示，G0/G1 期细胞群体增加，而 G2/M 期细胞减少。治疗前，与 HSC 相比，MPNST 细胞中 Axl、Tyro3 和乙酰化 H3 的表达明显升高。用 Calebin-A 处理后，磷酸化 AKT、ERK1/2、存活素、端粒酶逆转录酶（hTERT）和乙酰化 H3 蛋白的表达减少。CHIP 检测显示，治疗后存活素（BRIC5）和 hTERT 基因的启动子 DNA 拷贝显著减少。HAT 的酶活性显著降低，但 HDAC 的酶活性没有降低。两种 HAT 抑制剂——表没食子儿茶素-3-没食子酸酯（EGCG）和漆树酸（AA）也对 MPNST 细胞表现出显著的抑制作用。最后，肿瘤大小的测量显示，用 Calebin-A 处理后，异种移植肿瘤的体积明显减小。
结论
体内外研究均表明，Calebin-A 可抑制 MPNST 的增殖，抑制存活素和 hTERT 的表达。这两个因子的表达减少可能是由于 HAT 活性降低导致组蛋白表观遗传修饰所致。
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