ERO1? inhibits cell apoptosis and regulates steroidogenesis in mouse granulosa cells
ERO1? inhibits cell apoptosis and regulates steroidogenesis in mouse granulosa cells
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ERO1α 抑制细胞凋亡并调节小鼠颗粒细胞的类固醇生成
DOI：
10.1016/j.mce.2020.110842
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发表时间：
2020-07-01
影响因子：
4.1
通讯作者：
Chen, Fenglei
中科院分区：
文献类型：
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作者：
Hu, Jiahui;Jin, Jiaqi;Chen, Fenglei
ER氧化还原蛋白1α（ERO 1 α）是存在于ER中的一种氧化酶，参与蛋白质的折叠和分泌，抑制细胞凋亡，调节肿瘤的进展，是一种新的肿瘤预后不良因子。然而，ERO 1 α的其他生理功能尚未被发现。虽然我们的初步研究结果表明ERO 1 α在卵巢中，特别是在颗粒细胞中有强表达，但ERO 1 α在卵泡发育中的作用尚不清楚。本研究旨在探讨ERO 1 α在卵巢颗粒细胞凋亡和类固醇合成中的作用及其可能机制。免疫组化结果显示，ERO 1 α主要定位于卵巢颗粒细胞和卵母细胞。Western blot分析表明，ERO 1 α在动情周期中动情期表达最高。通过将ERO 1 α过表达和敲低慢病毒转导入原代培养的颗粒细胞，检测ERO 1 α对细胞凋亡和类固醇合成的影响。流式细胞仪分析显示ERO 1 α可减少颗粒细胞凋亡。Western blot和RT-qPCR分析发现，ERO 1 α使BCL-2/BAX比值升高，BAD和Caspase-3表达降低。ELISA分析显示ERO 1 α可促进雌激素（E2）的分泌。Western blot和RT-qPCR分析发现，ERO 1 α可增加星星、CYP 11 A1、3β-HSD、CYP 17 A1和CYP 19 A1的表达，降低CYP 1B 1的表达。Western blot分析发现，ERO 1 α通过增加AKT和P70 S6激酶的磷酸化水平，激活PI 3 K/AKT/mTOR信号通路，增加PDI和PRDX 4的表达。总之，这些结果表明，ERO 1 α可能发挥抗凋亡作用，并调节颗粒细胞中的类固醇合成，至少部分，通过激活PI 3 K/AKT/mTOR信号通路。
ER oxidoreduclin 1α (ERO1α), an oxidase that exists in the ER, participates in protein folding and secretion and inhibiting apoptosis, and regulates tumor progression, which is a novel factor of poor cancer prognosis. However, the other physiological functions of ERO1α remain undiscovered. Although our preliminary results of this study indicated that ERO1α revealed the robust expression in ovary, especially in granulosa cells, the role of ERO1α in follicular development is not well known. Therefore, the aims of the present study were to explore the role of ERO1α and the possible mechanisms in regulating cell apoptosis and steroidogenesis in ovarian granulosa cells. ERO1α was mainly localized in granulosa cells and oocytes in the adult ovary by immunohistochemistry. Western blot analysis showed that the expression of ERO1α was highest at oestrous stage during the estrous cycle. The effect of ERO1α on cell apoptosis and steroidogenesis was detected by transduction of ERO1α overexpression and knockdown lentiviruses into primary cultured granulosa cells. Flow cytometry analysis showed that ERO1α decreased granulosa cells apoptosis. Western bolt and RT-qPCR analysis found that ERO1α increased the ratio of BCL-2/BAX, and decreased BAD and Caspase-3 expression. ELISA analysis showed that ERO1α enhanced estrogen (E2) secretion. Western bolt and RT-qPCR analysis found that ERO1α increased StAR, CYP11A1, 3β-HSD, CYP17A1, and CYP19A1 expression, and decreased CYP1B1 expression. Furthermore, Western bolt analysis found that ERO1αincreased PDI and PRDX 4 expression, and activated the PI3K/AKT/mTOR signaling pathway through increasing the phosphorylation of AKT and P70 S6 kinase. In summary, these results suggested that ERO1α might play an anti-apoptotic role and regulate steroidogenesis in granulosa cells, at least partly, via activation of the PI3K/AKT/mTOR signaling pathway.