Knockdown of histone H1–5 gene affects the sensitivity of MRC-5 and HeLa cells to DNA damaging agents

IF 0.7 Q4 GENETICS & HEREDITY

Human Gene Pub Date : 2025-09-12 DOI:10.1016/j.humgen.2025.201475

Tigran Harutyunyan , Anzhela Sargsyan , Gohar Tadevosyan , Lily Kalashyan , Rouben Aroutiounian , Galina Hovhannisyan

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Abstract

Linker histone H1.5, encoded by the H1–5 gene, plays a key role in chromatin compaction and genome stability. However, its role in cellular responses to mutagens is still poorly understood. Here, we analyzed the genotoxic effects of the standard genotoxic drugs doxorubicin (DOX) and mitomycin C (MMC) at non-cytotoxic concentrations after 24 h of treatment in normal lung fibroblasts (MRC-5) and cervical carcinoma cells (HeLa) with H1–5 gene knockdown (KD). Using CRISPR-Cas9, we achieved significant repression of H1–5 expression. Alkaline DNA comet assay and cytokinesis-block micronucleus assay showed that H1–5 KD significantly increased spontaneous and mutagen-induced DNA and chromosome damage levels in both cell lines (p < 0.05). Our results suggest that H1–5 gene deficiency makes DNA more susceptible to genotoxic impact while its mechanistic role in occurrence of DNA and chromosome damage requires further elucidation.

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组蛋白H1-5基因敲低影响MRC-5和HeLa细胞对DNA损伤剂的敏感性

由H1-5基因编码的连接蛋白H1.5在染色质压实和基因组稳定性中起关键作用。然而，它在细胞对诱变剂反应中的作用仍然知之甚少。在这里，我们分析了标准基因毒性药物多柔比星（DOX）和丝裂霉素C （MMC）在治疗24小时后的非细胞毒性浓度对正常肺成纤维细胞（MRC-5）和H1-5基因敲低（KD）的宫颈癌细胞（HeLa）的遗传毒性作用。使用CRISPR-Cas9，我们实现了H1-5表达的显著抑制。碱性DNA彗星试验和细胞分裂阻滞微核试验显示，H1-5 KD显著增加了两种细胞系的自发和诱变诱导的DNA和染色体损伤水平（p < 0.05）。我们的研究结果表明，H1-5基因缺失使DNA更容易受到遗传毒性的影响，但其在DNA和染色体损伤发生中的机制有待进一步阐明。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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