The association between high mobility group box 1 chromatin protein and mitotic chromosomes in glioma cells
The association between high mobility group box 1 chromatin protein and mitotic chromosomes in glioma cells
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DOI：
10.3892/ol.2019.11170
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发表时间：
2020-01-01
期刊：
影响因子：
2.9
通讯作者：
Yao, Anhui
中科院分区：
文献类型：
--
作者：
Jia, Liyun;Song, Huiling;Yao, Anhui
高迁移率族蛋白 1 (HMGB1) 是一种丰富的非组蛋白核蛋白，作为染色质的结构蛋白，调节基因组复制和重组、mRNA 转录和 DNA 修复。 HMGB1 与多种癌症类型的肿瘤发生有关，并且在神经胶质瘤细胞中已证实 HMGB1 上调。然而，HMGB1 与神经胶质瘤有丝分裂染色体之间的关联仍不清楚。在本研究中，研究了 HMGB1 在神经胶质瘤组织和细胞中的亚细胞定位。此外，使用人类 HMGB1 蛋白的增强型绿色荧光蛋白 (EGFP) 标记和染色体扩散来研究 HMGB1 与有丝分裂染色体的结合。目前的研究结果表明，HMGB1定位于细胞核和细胞质，并确定其与多聚甲醛（PFA）固定的胶质瘤组织中增殖细胞的浓缩染色体结合。然而，在体外用冷冻甲醇和 5% (v/v) 乙酸或 PFA 固定时，HMGB1 也与间期相关（但与有丝分裂染色体无关）。来自活细胞成像和染色体扩散的数据表明 HMGB1 与神经胶质瘤细胞中有丝分裂染色体的关联。目前的结果表明，HMGB1 与神经胶质瘤细胞中的有丝分裂染色体结合，并且固定剂的使用可能会导致 HMGB1-DNA 相互作用的解离。因此，在活体标本和染色体扩散中，EGFP融合蛋白可能是确定胶质瘤细胞中HMGB1正确定位和相互作用的准确指标。
High mobility group box 1 (HMGB1) is an abundant non-histone nuclear protein that functions as a structural protein of chromatin, regulating genome replication and recombination, mRNA transcription and DNA repair. HMGB1 has been implicated in the tumorigenesis of various cancer types, and the upregulation of HMGB1 has been demonstrated in glioma cells. However, the association between HMGB1 and the mitotic chromosomes in glioma remains uncharacterized. In the present study, the sub-cellular localization of HMGB1 in glioma tissues and cells was investigated. In addition, enhanced green fluorescent protein (EGFP)-tagging of the human HMGB1 protein and chromosome spreading were used to investigate the combination of HMGB1 with mitotic chromosomes. The results of the current study indicated that HMGB1 was localized to the nucleus and the cytoplasm, and it was determined to combine with the condensed chromosomes of proliferating cells in paraformaldehyde (PFA)-fixed glioma tissues. However, HMGB1 was also associated with interphase (but not mitotic chromosomes) when fixed with chilled methanol and 5% (v/v) acetic acid or PFA in vitro. Data from live cell imaging and chromosome spreading indicated the association of HMGB1 with mitotic chromosomes in glioma cells. The present results suggest that HMGB1 combines with mitotic chromosomes in glioma cells, and that the use of fixatives may result in the dissociation of the HMGB1-DNA interaction. Therefore, in live specimens and chromosome spreads, EGFP fusion proteins may represent an accurate indicator for the determination of the correct localization and interaction of HMGB1 in glioma cells.