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.

中文翻译：

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ERO1α抑制细胞凋亡并调节小鼠颗粒细胞中的类固醇生成。

ER氧化还原蛋白1α（ERO1α）是ER中存在的一种氧化酶，参与蛋白质折叠和分泌并抑制细胞凋亡，并调节肿瘤的进展，这是癌症预后不良的新因素。但是，ERO1α的其他生理功能仍未发现。尽管我们这项研究的初步结果表明ERO1α在卵巢中，尤其是在颗粒细胞中显示出了强大的表达，但ERO1α在卵泡发育中的作用尚不清楚。因此，本研究的目的是探讨ERO1α的作用及其在调节卵巢颗粒细胞中细胞凋亡和类固醇生成中的可能机制。ERO1α主要通过免疫组织化学定位在成年卵巢的颗粒细胞和卵母细胞中。Western印迹分析表明，在动情周期中，ERO1α的表达在发情期最高。通过将ERO1α过表达和击倒慢病毒转导至原代培养的颗粒细胞中，检测到ERO1α对细胞凋亡和类固醇生成的影响。流式细胞仪分析表明ERO1α降低了颗粒细胞的凋亡。Western螺栓和RT-qPCR分析发现ERO1α增加BCL-2 / BAX的比率，并降低BAD和Caspase-3的表达。ELISA分析表明ERO1α增强了雌激素（E2）的分泌。Western bolt和RT-qPCR分析发现ERO1α增加StAR，CYP11A1、3β-HSD，CYP17A1和CYP19A1表达，并降低CYP1B1表达。此外，Western bolt分析发现ERO1α增加了PDI和PRDX 4的表达，并通过增加AKT和P70 S6激酶的磷酸化来激活PI3K / AKT / mTOR信号通路。总之，这些结果表明，ERO1α可能至少部分通过激活PI3K / AKT / mTOR信号通路来发挥抗凋亡作用，并调节颗粒细胞中的类固醇生成。