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IER5在γ射线照射后对HeLa细胞中Cdc25B的表达起负调控作用。

IER5 Negatively Regulates Cdc25B Expression in HeLa Cells After Gamma Ray Irradiation.

作者信息

Zhao Xianzhe, Ding Lixin, Ma Yongzhong, Cheng Wei, Zhang Li, Ding Kuke

机构信息

The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.

National Institute for Radiological Protection, China CDC, Beijing, China.

出版信息

Biomed Res Int. 2025 Aug 14;2025:5644318. doi: 10.1155/bmri/5644318. eCollection 2025.

Abstract

Radiotherapy is commonly used to treat many cancers, and their sensitivity to radiation is crucial for favorable outcomes. This study investigated whether the immediate early response 5 (IER5) protein affects Cdc25B protein expression in HeLa cells after gamma irradiation. IER5 was knocked down using RNA interference (RNAi) in HeLa cells irradiated with 2 or 4 Gy of gamma rays. The mRNA and protein expression levels were subsequently determined via qRT-PCR and western blotting. The distribution of cells during the cell cycle was also determined using flow cytometry. IER5 protein levels were successfully reduced with RNAi. Variations in IER5 levels led to differences in Cdc25B mRNA and protein levels. Moreover, IER5 affected the proportion of cells in the G2 phase, which is regulated mainly by Cdc25B. Pearson correlation analysis was conducted on the expression levels of IER5 and Cdc25B in HeLa cells and the IER5-silenced HeLa (siIER5-HeLa) cell line at various time points after exposure to 4 Gy of gamma rays, and a negative correlation was detected between IER5 and Cdc25B expression levels, with correlation coefficients of -0.686 and -0.663, respectively. Additionally, variations in IER5 levels led to differences in the expression levels of p53, NF-YB, and p300, which may be putative transcriptional regulators of Cdc25B. These results suggest that IER5 plays a negative role in regulating Cdc25B expression, which may involve interactions with the transcriptional regulators p53, NF-YB, and p300.

摘要

放射疗法常用于治疗多种癌症,而它们对辐射的敏感性对于取得良好疗效至关重要。本研究调查了即刻早期反应5(IER5)蛋白是否会影响γ射线照射后HeLa细胞中Cdc25B蛋白的表达。在用2或4 Gyγ射线照射的HeLa细胞中,使用RNA干扰(RNAi)敲低IER5。随后通过qRT-PCR和蛋白质印迹法测定mRNA和蛋白质表达水平。还使用流式细胞术确定细胞周期中细胞的分布。RNAi成功降低了IER5蛋白水平。IER5水平的变化导致Cdc25B mRNA和蛋白质水平的差异。此外,IER5影响G2期细胞的比例,而G2期主要由Cdc25B调节。对HeLa细胞和IER5沉默的HeLa(siIER5-HeLa)细胞系在暴露于4 Gyγ射线后的不同时间点的IER5和Cdc25B表达水平进行Pearson相关性分析,检测到IER5和Cdc25B表达水平之间呈负相关,相关系数分别为-0.686和-0.663。此外,IER5水平的变化导致p53、NF-YB和p300表达水平的差异,它们可能是Cdc25B的假定转录调节因子。这些结果表明,IER5在调节Cdc25B表达中起负作用,这可能涉及与转录调节因子p53、NF-YB和p300的相互作用。

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