抗癌植物次生代谢产物从染色质中诱导连接组蛋白耗竭。

Anticancer Plant Secondary Metabolites Induce Linker Histone Depletion from Chromatin.

机构信息

N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia, 115478 Moscow, Russia.

Federal State Budgetary Institution, Centre for Strategic Planning and Management of Biomedical Health Risks of the Federal Medical and Biological Agency, 119121 Moscow, Russia.

出版信息

Front Biosci (Landmark Ed). 2024 Aug 5;29(8):275. doi: 10.31083/j.fbl2908275.

DOI:10.31083/j.fbl2908275

PMID:39206902

Abstract

BACKGROUND

Many plant secondary metabolites (PSMs) were shown to intercalate into DNA helix or interact with DNA grooves. This may influence histone-DNA interactions changeing chromatin structure and genome functioning.

METHODS

Nucleosome stability and linker histone H1.2, H1.4 and H1.5 localizations were studied in HeLa cells after the treatment with 15 PSMs, which are DNA-binders and possess anticancer activity according to published data. Chromatin remodeler CBL0137 was used as a control. Effects of PSMs were studied using fluorescent microscopy, flowcytometry, quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), western-blotting.

RESULTS

We showed that 1-hour treatment with CBL0137 strongly inhibited DNA synthesis and caused intensive linker histone depletion consistent with nucleosome destabilization. None of PSMs caused nucleosome destabilization, while most of them demonstrated significant influence on linker histone localizations. In particular, cell treatment with 11 PSMs at non-toxic concentrations induced significant translocation of the histone H1.5 to nucleoli and most of PSMs caused depletion of the histones H1.2 and H1.4 from chromatin fraction. Curcumin, resveratrol, berberine, naringenin, and quercetin caused significant redistribution of all three variants of the studied linker histones showing some overlap of PSM effects on linker histone DNA-binding. We demonstrated that PSMs, which induced the most significant redistribution of the histone H1.5 (berberine, curcumin and naringenin), influence the proportion of cells synthesizing DNA, expressing or non-expressing cyclin B and influence cell cycle distribution. Berberine induction of H1.5 translocations to nucleoli was shown to occur independently on the phases of cell cycle (metaphase was not analyzed).

CONCLUSIONS

For the first time we revealed PSM influence on linker histone location in cell nuclei that opens a new direction of PSM research as anticancer agents.

摘要

背景

许多植物次生代谢产物（PSMs）被证明可以嵌入 DNA 双螺旋或与 DNA 凹槽相互作用。这可能会影响组蛋白-DNA 相互作用，改变染色质结构和基因组功能。

方法

根据已发表的数据，我们研究了在 HeLa 细胞中，15 种具有 DNA 结合活性和抗癌活性的 PSM 处理后核小体稳定性和连接组蛋白 H1.2、H1.4 和 H1.5 的定位。使用染色质重塑剂 CBL0137 作为对照。使用荧光显微镜、流式细胞术、实时定量聚合酶链反应（RT-qPCR）和蛋白质印迹法研究 PSM 的作用。

结果

我们表明，1 小时的 CBL0137 处理强烈抑制 DNA 合成，并导致连接组蛋白严重耗竭，与核小体不稳定一致。没有一种 PSM 导致核小体不稳定，但大多数 PSM 对连接组蛋白的定位有显著影响。特别是，在非毒性浓度下，用 11 种 PSM 处理细胞会导致组蛋白 H1.5 向核仁的显著易位，并且大多数 PSM 会导致染色质部分的组蛋白 H1.2 和 H1.4 耗竭。姜黄素、白藜芦醇、小檗碱、柚皮素和槲皮素导致研究连接组蛋白的所有三种变体的显著再分布，显示 PSM 对连接组蛋白 DNA 结合的影响有一些重叠。我们证明，诱导组蛋白 H1.5 重分布最显著的 PSM（小檗碱、姜黄素和柚皮素）会影响合成 DNA、表达或不表达细胞周期蛋白 B 的细胞比例，并影响细胞周期分布。小檗碱诱导 H1.5 向核仁易位的发生独立于细胞周期的阶段（未分析有丝分裂期）。