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选择稳定基因参考基因用于激活和非激活PBMC的基因表达研究在正常和缺氧条件下

Artur Wardaszka¹, Anna Smolarska², Piotr Bednarczyk¹

¹Department of Physics and Biophysics, Institute of Biology, Warsaw University of Life Sciences, 02-787 Warsaw, Poland.

International journal of molecular sciences

|July 29, 2025

概括

这项研究确定了稳定的参考基因,用于分析缺氧下免疫细胞基因表达,这对于了解癌症免疫疗法的有效性至关重要. 建议使用RPL13A,S18和SDHA,以在低氧条件下获得准确的结果.

科学领域:

免疫学免疫学免疫学
分子生物学分子生物学
癌症研究癌症研究

背景情况:

免疫疗法在癌症中的有效性因瘤微环境因素如缺氧而有所不同.
缺氧会损害免疫细胞的功能,影响基因表达,需要进行精确的分析.
在低氧条件下,人类外周血液单核细胞中参考基因稳定性的数据有限.

研究的目的:

在各种低氧条件下,评估常用参考基因在人体外周血液单核细胞 (PBMCs) 中的表达稳定性.
在涉及缺氧的免疫学研究中确定可靠的基因参考基因,用于准确的基因表达分析.
为未来研究低氧对癌症免疫疗法中免疫细胞的影响提供关键数据.

主要方法:

在PBMC中评估了七个参考基因 (S18,HPRT,IPO8,RPL13A,SDHA,PPIA,UBE2D2) 的稳定性.
在24小时的时间内,细胞在正常性,缺氧性 (1% O2) 和化学诱导的缺氧条件下进行培养.
使用delta Ct,geNorm,NormFinder和BestKeeper算法分析了基因表达的稳定性.

主要成果:

在低氧条件下,RPL13A,S18和SDHA被确定为最稳定的参考基因.
IPO8和PPIA表现最不稳定,因此不适合作为参考.
建立了可靠的基因参考基因,用于对暴露于缺氧的人类PBMC的基因表达分析.

结论:

准确的基因基因选择对于免疫学研究中可靠的基因表达分析至关重要,特别是在缺氧条件下.
确定了稳定的参考基因 (RPL13A,S18,SDHA) 将加强对癌症中缺氧驱动的免疫细胞重编程的研究.
这项研究通过提供分子分析的基本工具,支持更好地了解癌症免疫治疗中的T细胞反应.

关键词:

这就是PBMCs.家庭管理基因的基因缺氧缺氧是指缺氧的情况.定量实时PCR (qRT-PCR) 是一种方法.基因基因基因基因基因基因基因基因基因基因

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