17β-estradiol suppresses H2O2-induced senescence in human umbilical vein endothelial cells by inducing autophagy through the PVT1/miR-31/SIRT3 axis

作者：Xiang, Xiuting; Wang, Yuyan; Huang, Guanshen; Huang, Jianming; Gao, Mingjian; Sun, Meihua; Xia, Hao; Pare, Rahmawati; Li, Jingjun^*; Ruan, Yunjun^*

来源：Journal of Steroid Biochemistry and Molecular Biology, 2023, 227: 106244.

DOI：10.1016/j.jsbmb.2022.106244

• 17?? -estradiol
• Autophagy
• Senescence
• PVT1
• miR-31
• SIRT3

Objective: 178-estradiol (178-E2) has been implicated in activating autophagy by upregulating SIRT3 (Sirtuin 3) expression, thereby inhibiting the senescence of vascular endothelial cells. Herein, we further examined the molecular mechanisms that regulate SIRT3 expression in 178-E2-induced autophagy.Methods: Reverse-transcription-polymerase chain reaction was employed to measure the expression of plasma-cytoma variant translocation 1 (PVT1), microRNAs (miRNAs), and SIRT3, and the dual-luciferase assay was used to determine their interaction. Electron microscopy observes autophagosomes, green fluorescent protein-microtubule-associated protein 1 light chain 3 (GFP-LC3) staining, and immunoblot analysis with antibodies against LC3,beclin-1, and P62 were conducted to measure autophagy. Cellular senescence was determined using immunoblot analysis with anti-phosphorylated retinoblastoma and senescence-associated 8-galactosidase staining.Results: Women with higher estrogen levels (during the 10-13th day of the menstrual cycle or premenopausal) exhibit markedly higher serum levels of PVT1 than women with lower estrogen levels (during the menstrual period or postmenopausal). The dual-luciferase assay showed that PVT1 acts as a sponge for miR-31, and miR-31 binds to its target gene, SIRT3. The 178-E2 treatment increased the expression of PVT1 and SIRT3 and down -regulated miR-31 expression in human umbilical vein endothelial cells (HUVECs). Consistently, PVT1 over -expression suppresses miR-31 expression, promotes 178-E2-induced autophagy, and inhibits H2O2-induced senescence. miR-31 inhibitor increases SIRT3 expression and leads to activation of 178-E2-induced autophagy and suppression of H2O2-induced senescence.Conclusion: Our findings demonstrated that 178-E2 upregulates PVT1 gene expression and PVT1 functions as a sponge to inhibit miR-31, resulting in the upregulation of SIRT3 expression and activation of autophagy and subsequent inhibition of H2O2-induced senescence in HUVECs.

• 单位
  南方医科大学