Downregulation of MCM2 contributes to the reduced growth potential of epithelial progenitor cells in chronic nasal inflammation
Downregulation of MCM2 contributes to the reduced growth potential of epithelial progenitor cells in chronic nasal inflammation
复制标题
MCM2 的下调导致慢性鼻炎症中上皮祖细胞的生长潜力降低
DOI：
10.1016/j.jaci.2020.11.026
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发表时间：
2021-05-05
影响因子：
14.2
通讯作者：
Li, Chun Wei
中科院分区：
文献类型：
--
作者：
Li, Li Yue;Zhou, Yu Tao;Li, Chun Wei
背景：最近的研究表明，人鼻上皮祖细胞（hNEPCs）在慢性鼻腔炎症中具有增殖能力差的特点。目的：我们试图研究慢性炎症鼻黏膜中hNEPCs生长潜力降低的关键分子功能和候选分子。方法：选取28例鼻息肉（NPs）患者和13例正常人的鼻活检标本。连续培养3代鼻标本hNEPCs，通过RNA测序分析其分子和功能谱。通过细胞周期、定量PCR和Western blot分析，在NPs患者和对照组的hNEPCs和组织样本中验证了微染色体维持蛋白（MCM）家族基因MCM2；小干扰rna介导的敲低试验；免疫荧光染色。结果：与对照hNEPCs相比，np衍生的hNEPCs表现出：(1)生长动力学降低，体现在菌落形成效率和倍增时间上；(2)抑制细胞周期进程，通过基因本体和/或通路和细胞周期分析证明；(3)下调MCM复合物的关键蛋白MCM2的表达，MCM2在G1/S检查点对DNA复制至关重要。此外，MCM2敲低的hNEPCs增殖率降低，异常重塑鼻上皮基底细胞中MCM2蛋白水平明显低于正常上皮。结论：这些结果表明NP组织的鼻基底上皮细胞和鼻祖上皮细胞的细胞周期进程受到抑制，MCM2下调，这可能是慢性炎症上呼吸道hNEPCs生长潜力降低的原因。[J]过敏症临床免疫学杂志，2011;47:366 - 366。
Background: Recent studies have shown that human nasal epithelial progenitor cells (hNEPCs) are characterized by poor proliferation capacities during chronic nasal inflammation. Objective: We sought to investigate the key molecular functions and candidates that contribute to the reduced growth potential of hNEPCs in chronically inflamed nasal mucosa.Methods: Nasal biopsy specimens were obtained from 28 patients with nasal polyps (NPs) and 13 healthy controls. hNEPCs from nasal samples were cultured for 3 consecutive passages, and their molecular and functional profiles were analyzed by RNA sequencing. The minichromosome maintenance protein (MCM) family gene MCM2 was validated in hNEPCs and tissue samples from patients with NPs and control subjects by cell cycle, quantitative PCR, and Western blot analyses; small interfering RNA-mediated knockdown assay; and immunofluorescent staining.Results: Compared with control hNEPCs, NP-derived hNEPCs showed (1) reduced growth kinetics, as evidenced by the colony forming efficiency and doubling time; (2) inhibited cell cycle progression, as evidenced by gene ontology and/or pathway and cell cycle analyses; and (3) downregulated expression of MCM2, the key protein of the MCM complex, which is critical for DNA replication at the G1/S checkpoint. Moreover, hNEPCs with MCM2 knockdown showed a decreased proliferation rate, and the MCM2 protein level in basal cells was significantly lower in abnormally remodeled nasal epithelium than in normal epithelium.Conclusion: These results demonstrate inhibited cell cycle progression and MCM2 downregulation in basal or progenitor nasal epithelial cells from NP tissue, which may contribute to the decreased growth potential of hNEPCs in chronically inflamed upper airways. (J Allergy Clin Immunol 2021;147:1966-73.)