生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (5): 11-.doi: 10.3969/j.issn.2095-1736.2023.05.011

• 生物医学专题 • 上一篇    下一篇

线粒体蛋白HIGD1A调控细胞自噬增强HeLa细胞辐射抗性

赵喜朋1,2， 赵国平2   

  1. 1. 安徽大学 物质科学与信息技术研究院， 合肥 230601；
    2. 中国科学院合肥物质科学研究院强磁场科学中心， 合肥 230031
  • 出版日期:2023-10-18 发布日期:2023-10-17
  • 通讯作者: 赵国平，博士，研究员，研究方向为生物物理学，E-mail:gpz@ipp.ac.cn
  • 作者简介:赵喜朋，硕士研究生，研究方向为生物物理学，E-mail:q20201134@stu.ahu.edu.cn
  • 基金资助:
    安徽省重点研发计划项目(202104a07020006)

HIGD1A regulates autophagy to enhance the radiation resistance of HeLa cells

ZHAO Xipeng1,2, ZHAO Guoping2   

  1. 1. Institutes of Physical Science and Information Technology, Anhui University, Hefei 230601, China; 2. Hefei
    Institutes of Physical Science Chinese Academy of Sciences High Magnetic Field Laboratory, Hefei 230031, China
  • Online:2023-10-18 Published:2023-10-17

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摘要： 为探究线粒体蛋白HIGD1A通过调控自噬增强宫颈癌细胞（HeLa）辐射敏感性的机制，以对照细胞和HIGD1A敲低的HeLa细胞为研究对象，利用自噬诱导剂Earle’s 平衡盐溶液（EBSS）和电离辐射处理细胞，Western Blot检测细胞自噬标志蛋白LC3、P62的表达水平，并检测Ad-mCherry-GFP-LC3B重组腺病毒感染细胞的荧光表达水平；CCK-8检测氯喹/雷帕霉素预处理2 h，并在辐照处理之后48 h的细胞活力；Western Blot检测雷帕霉素处理之后的caspase-7和cleaved caspase-3蛋白表达水平。结果显示：与对照细胞相比，敲低HIGD1A抑制EBSS和电离辐射处理之后LC3的生成、P62的降解和mCherry-GFP-LC3黄色斑点、红色斑点的生成；CCK-8检测结果发现，敲低HIGD1A并结合氯喹处理降低了细胞在电离辐射处理之后的细胞活力；此外，敲低HIGD1A显著增加了肿瘤细胞中caspase-7和cleaved caspase-3蛋白表达水平，在雷帕霉素处理后HIGD1A敲低细胞中仍存在较高的凋亡水平。结果表明，HIGD1A通过自噬途径参与调节肿瘤细胞的辐射敏感性。

关键词: HIGD1A, 细胞凋亡, 细胞自噬, 细胞活力, 辐射敏感性

Abstract: This article mainly explored the mechanism of mitochondrial protein HIGD1A enhancing the radiosensitivity of cervical cancer cells (HeLa) by regulating autophagy. Using control cells and HIGD1A knockdown HeLa cells as research subjects, cells were treated with autophagy inducer Earle’s balanced salt solution (EBSS) and ionizing radiation. The expression levels of cell autophagy marker proteins LC3 and P62 were detected by Western Blot, and the fluorescence expression level of Ad-mCherry-GFP-LC3B recombinant adenovirus infected cells was detected. Cells were irradiated after the pretreatment of chloroquine/rapamycin for 2 hours, then their viabilities were detected using CCK-8 48 hours later. The expression levels of caspase-7 and cleaved caspase-3 proteins were detected by Western Blot after rapamycin treatment. The results showed that compared with the control cells, knocking down HIGD1A inhibited the generation of LC3, P62 degradation, and the generation of yellow and red spots in mCherry-GFP-LC3B after EBSS or ionizing radiation treatment. CCK-8 assay results showed that knockdown of HIGD1A combined with chloroquine treatment reduced cell viability after ionizing radiation treatment. In addition, knocking down HIGD1A significantly increased the expression levels of caspase-7 and cleaved caspase-3 proteins in tumor cells, and there were still high levels of apoptosis in HIGD1A knocked down cells after rapamycin treatment. The results indicated that HIGD1A was involved in regulating the radiation sensitivity of tumor cells through the autophagy pathway.

Key words: HIGD1A, apoptosis, autophagy, cell viability, radiosensitivity

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