| 文映人,黄进,杨雨馨,等.HIST2H2BE乳酸化参与调控乳腺癌细胞辐射抗性机制的研究[J].中华放射医学与防护杂志,2026,46(4):329-335.Wen Yingren,Huang Jin,Yang Yuxin,et al.Mechanisms underlying HIST2H2BE lactylation in regulating radiation resistance in breast cancer cells[J].Chin J Radiol Med Prot,2026,46(4):329-335 |
| HIST2H2BE乳酸化参与调控乳腺癌细胞辐射抗性机制的研究 |
| Mechanisms underlying HIST2H2BE lactylation in regulating radiation resistance in breast cancer cells |
| 投稿时间：2025-10-20 |
| DOI：10.3760/cma.j.cn112271-20251020-00365 |
| 中文关键词: 乳腺癌|乳酸化|辐射抵抗|HIST2H2BE |
| 英文关键词:Breast cancer|Lactylation|Radiation resistance|HIST2H2BE |
| 基金项目:国家自然科学基金(82574034);浙江省自然科学基金(LMS25H220002) |
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| 中文摘要: |
| 目的 探究组蛋白H2B变体HIST2H2BE的乳酸化修饰在调控乳腺癌细胞辐射抗性中的作用及其分子机制。方法 通过分析Gene Expression Omnibus (GEO)数据库中乳腺癌患者放疗前后的转录组数据集GSE59733,筛选差异表达基因并与乳酸化相关基因取交集。利用乳腺癌细胞系MCF-7进行实验,通过Western blot检测电离辐射后乳腺癌细胞整体乳酸化水平及HIST2H2BE特异性乳酸化水平的变化;使用在线工具DeepKla预测潜在乳酸化位点,并通过构建赖氨酸-精氨酸点突变体质粒(K31R、K44R、K35R),结合免疫共沉淀技术验证关键修饰位点。采用CCK-8法和流式细胞术分别检测细胞活力与死亡率,评估HIST2H2BE及其K35位点乳酸化对细胞辐射敏感性的影响。结果 HIST2H2BE在乳腺癌组织及细胞中高表达。电离辐射可诱导乳腺癌细胞整体乳酸化水平升高,并特异性促进HIST2H2BE发生乳酸化修饰,其中K35为其关键乳酸化位点。功能实验表明,将其K35位点突变(K35R)以消除乳酸化修饰后,乳腺癌细胞活力降低(t = 7.57、8.91,P < 0.05),辐射诱导的细胞死亡率显著升高,表明细胞的辐射敏感性增强(t = 20.15、49.46,P < 0.05)。结论 HIST2H2BE在K35位点的乳酸化修饰是乳腺癌细胞获得辐射抗性的重要机制之一。靶向抑制HIST2H2BE K35乳酸化可能成为逆转乳腺癌放疗抵抗的潜在策略。 |
| 英文摘要: |
| Objective To investigate the role of the lactylation of histone H2B variant (HIST2H2BE) in regulating radiation resistance in breast cancer cells, along with the underlying mechanisms. Methods Differentially expressed genes (DEGs) were selected by analyzing the transcriptomic dataset GSE59733, which included pre- and post-radiotherapy data from breast cancer patients in the GEO database. Then, the intersection of DEGs and lactylation-related genes was determined. Based on experiments on the MCF-7 breast cancer cell line, changes in both overall lactylation levels of breast cancer cells and HIST2H2BE-specific lactylation were detected after ionizing radiation using Western blot. Potential lactylation sites were predicted using online tool DeepKla. Subsequently, key sites were validated using co-immunoprecipitation by constructing plasmids K31R, K44R, and K35R encoding lysine-to-arginine mutations. Cell viability and mortality were assessed using the CCK-8 and flow cytometry, respectively, to assess the effects of HIST2H2BE and its lactylation at site K35 on cellular radiosensitivity. Results HIST2H2BE exhibited high expression in breast cancer tissues and cells. Ionizing radiation induced both increased global lactylation levels of breast cancer cells and the specific promotion of HIST2H2BE lactylation, with K35 identified as the key lactylation site. Cell function experiments indicate that lactylation elimination by K35-to-K35R mutation led to reduced breast cancer cell viability (t =7.57, 8.91, P < 0.05) and significantly elevated radiation-induced cell mortality, indicating enhanced radiosensitivity (t =20.15, 49.46, P < 0.05) Conclusions The lactylation of HIST2H2BE at site K35 represents an important mechanism underlying radiation resistance in breast cancer cells. Therefore, targeted inhibition of such lactylation might serve as a potential strategy to reverse radiotherapy resistance in breast cancer. |
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