Ubiquitin-like modifier 1 ligating enzyme 1 relieves cisplatin-induced premature ovarian failure by reducing endoplasmic reticulum stress in granulosa cells

Abstract

Background Ubiquitin-like modifier 1 ligating enzyme 1 (UFL1), the ligase of the UFMylation system, has recently been reported to be involved in apoptosis and endoplasmic reticulum stress (ER stress) in a variety of diseases. Premature ovarian failure (POF) is a gynecological disease that severely reduces the fertility of women, especially in female cancer patients receiving chemotherapy drugs. Whether UFL1 is involved in protection against chemotherapy-induced POF and its mechanism remain unclear. Methods In this study, we examined the function of UFL1 in ovarian dysfunction and granulosa cell (GC) apoptosis induced by cisplatin through histological examination and cell viability analysis. We used western blotting, quantitative real-time PCR (qPCR) and immunofluorescence (IF) to detect the expression of UFL1 and the levels of ER stress specific markers. Enzyme linked immunosorbent assays were used to detect the levels of follicle-stimulating hormone (FSH) and estrogen (E2) in ovaries and GCs. In addition, we used infection with lentiviral particle suspensions to knock down and overexpress UFL1 in ovaries and GCs, respectively. Results Our data showed that the expression of UFL1 was reduced in POF model ovaries, accompanied by ER stress. In vitro, cisplatin induced a stress-related increase in UFL1 expression in GCs and enhanced ER stress, which was aggravated by UFL1 knockdown and alleviated by UFL1 overexpression. Furthermore, UFL1 knockdown resulted in a decrease in ovarian follicle number, an increase in atretic follicles, and decreased expression of AMH and FSHR. Conversely, the overexpression of UFL1 reduced cisplatin-induced damage to the ovary in vitro. Conclusions Our research indicated that UFL1 regulates cisplatin-induced ER stress and apoptosis in GCs, and participates in protection against cisplatin-induced POF, providing a potential therapeutic target for the clinical prevention of chemotherapeutic drug-induced POF. Graphical Abstract

Emerging role of UFMylation in secretory cells involved in the endocrine system by maintaining ER proteostasis

“…A mouse model of POF was set up by intraperitoneal injection of cisplatin. In this model, cell apoptosis and follicular atresia were triggered by elevated ER stress in GCs associated with a significant decrease of UFL1 protein expression there ( 79 ). Interestingly, overexpression of UFL1 could alleviate ER stress and reduce cisplatin-induced damage to the ovary, in the end augmenting follicle number and restoring estrogen secreted by GCs to some extent.…”

Premature ovarian insufficiency: a review on the role of oxidative stress and the application of antioxidants

“…Endogenous antioxidant systems exist within the body and cells, including antioxidant enzymes such as SOD, GPX, glutathione sulfhydryltransferase (GST), and CAT, in addition to GSH as a scavenger of ROS (205,223). Superoxide anions in ROS form a spontaneous conversion to H 2 O 2 catalyzed by SOD, and then CAT and GPX further convert H 2 O 2 to O 2 and H 2 O, which exerts antioxidant stress effects in POI patients (254). In addition, phase II detoxification enzymes have defensive effects against oxidative damage caused by exogenous chemical exposure, mainly including HO-1, HQO1 and g-glutamylcysteine ligase (g-GCL), which can moderate POI to some extent by counteracting OS.…”

“…HO-1 is a classical phase II detoxifying enzyme involved in preventing the generation of more active hydroxyl groups from H 2 O 2 , and also catalyzes the production of biliverdin from heme, of which the latter can form bilirubin, and biliverdin and bilirubin are powerful free radical scavengers in the body. It has been proved in mouse that g-GCLase catalyzes the formation of GSH and is the most critical ratelimiting enzyme in GSH biosynthesis (254). Furthermore, OS results in ER stress, and Ufm1-specific ligase 1 (UFL1) alleviates cisplatin-induced apoptosis in ovarian GCs by attenuating ER stress, thereby relieving POI to some extent (254).The key to POI is the reduction in the quantity and quality of oocytes, which results in symptoms such as hot flashes, excessive sweating, vaginal dryness, and even causes serious consequences such as infertility.…”

“…It has been proved in mouse that g-GCLase catalyzes the formation of GSH and is the most critical ratelimiting enzyme in GSH biosynthesis (254). Furthermore, OS results in ER stress, and Ufm1-specific ligase 1 (UFL1) alleviates cisplatin-induced apoptosis in ovarian GCs by attenuating ER stress, thereby relieving POI to some extent (254).The key to POI is the reduction in the quantity and quality of oocytes, which results in symptoms such as hot flashes, excessive sweating, vaginal dryness, and even causes serious consequences such as infertility.…”

Metformin protects ovarian granulosa cells in chemotherapy-induced premature ovarian failure mice through AMPK/PPAR-γ/SIRT1 pathway

“…According to the method of Tang and Shen [14,30], we mechanically isolated primary GCs from ovarian follicles and cultured them under appropriate conditions. The ovaries were collected from 3-4weeks mice, until 46-48 hours after the intraperitoneal injection of 20U pregnant mare serum gonadotropin (PMSG), we collected the ovaries and mechanically separated GCs from ovarian follicles aseptically.…”