Atg9a和Atg9b在生殖器官细胞中的不同表达模式。

IF 1.8 Q3 OBSTETRICS & GYNECOLOGY Clinical and Experimental Reproductive Medicine-CERM Pub Date : 2024-05-17 DOI:10.5653/cerm.2023.06737
Minseo Lee, Sujin Son, Hyunjung J Lim, Haengseok Song
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引用次数: 0

摘要

目的:自噬是一种主要的细胞内分解代谢途径,由自噬相关基因(Atg)编码的蛋白质依次发挥作用。ATG9 是参与这一过程的唯一跨膜蛋白,它在自噬的早期阶段调节磷脂向自噬体的转运。据报道,哺乳动物中有两种 Atg9 异构体:Atg9a和Atg9b。在这项研究中,我们考察了这两种异构体在小鼠体内的分子和细胞特征是否存在差异:方法:在小鼠妊娠第 1、4 和 8 天收集整个子宫,并对卵巢切除的小鼠注射药物、黄体酮或 17β-雌二醇。收集的细胞来自生殖组织,如颗粒细胞、子宫上皮细胞(UEC)、子宫基质细胞(USC)和卵母细胞。本分析还使用了两种人类子宫细胞系。研究人员进行了逆转录聚合酶链反应试验、免疫荧光染色。使用血清饥饿条件诱导原代细胞自噬:结果:Atg9a和Atg9b在多种小鼠组织和生殖细胞中表达。Atg9A和Atg9B对类固醇激素的反应均无明显变化。对UECs和USCs的免疫荧光染色显示,ATG9A呈点状分布,而ATG9B在细胞质中呈细长管状分布。在人类癌细胞系中,检测不到ATG9B,而在所有检测的细胞类型中都发现了ATG9A:结论:Atg9异构体在子宫内膜癌细胞中表现出不同的亚细胞定位,可能在自噬过程中发挥不同的作用。值得注意的是,人类子宫细胞的ATG9B表达量减少,这表明这种抑制可能是由于表观遗传调控所致。
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The differential expression patterns of Atg9a and Atg9b in cells of the reproductive organs.

Objective: Autophagy is a major intracellular catabolic pathway governed by the sequential actions of proteins encoded by autophagy-related genes (Atg). ATG9, the only transmembrane protein involved in this process, regulates phospholipid translocation to autophagosomes during the early phases of autophagy. In mammals, two Atg9 isoforms have been reported: Atg9a and Atg9b. In this study, we examined whether the molecular and cellular characteristics of these two isoforms differed in mice.

Methods: Whole uteri were collected on days 1, 4, and 8 of pregnancy and from ovariectomized mice injected with vehicle, progesterone, or 17β-estradiol. Cells from reproductive tissues, such as granulosa cells, uterine epithelial cells (UECs), uterine stromal cells (USCs), and oocytes were collected. Two human uterine cell lines were also used in this analysis. Reverse transcription-polymerase chain reaction tests, Western blotting, and immunofluorescence staining were performed. Serum starvation conditions were used to induce autophagy in primary cells.

Results: Atg9a and Atg9b were expressed in multiple mouse tissues and reproductive cells. Neither Atg9A nor Atg9B significantly changed in response to steroid hormones. Immunofluorescence staining of the UECs and USCs showed that ATG9A was distributed in a punctate-like pattern, whereas ATG9B exhibited a pattern of elongated tubular shapes in the cytoplasm. In human cancer cell lines, ATG9B was undetectable, whereas ATG9A was found in all cell types examined.

Conclusion: The Atg9 isoforms exhibited distinct subcellular localizations in UECs and may play different roles in autophagy. Notably, human uterine cells exhibited reduced ATG9B expression, suggesting that this suppression may be due to epigenetic regulation.

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