RESUMEN
Polycystic ovarian syndrome (PCOS) is a frequent metabolic disorder in premenopausal woman, featured with increased androgen, reduced ovulation and insulin resistance. An increasing number of reports have confirmed that microRNAs (miRNAs) play pivotal roles in PCOS. However, the diagnostic mechanisms of microRNA (miR)-206 in PCOS remain unclear. Liquiritin, extracted from Glycyrrhiza Radix, has multiple pharmacological activities in diseases, including PCOS. Our report was designed to explore whether liquiritin play a role in PCOS by regulating human ovarian granulosa cell-like KGN cells proliferation and apoptosis through miR-206/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway, and to further elucidate the underlying molecular mechanisms. KGN cells were exposed to various concentration (0, 20, 40, 80 µM) of liquiritin for 48 h, cell proliferation and apoptosis were measured by 3-(45)-dimethylthiahiazo(-z-y1)-35-di-phenytetrazoliumromide (MTT) and flow cytometry analysis. Reverse transcriptionquantitative polymerase chain reaction was conducted to checked the levels of miR-206 in KGN cells. The protein expression levels of cleaved caspase3, caspase3, phosphorylated (p)-AKT and AKT were analyzed using Western blot assay. We found that liquiritin stimulation led to reduced viability and enhanced apoptotic KGN cells, which along with increased cleaved caspase3 and cleaved caspase3/caspase3 ratio. Moreover, liquiritin obviously reduced p-AKT expression and p-AKT/AKT ratio. MiR-206 was up-regulated in liquiritin-treated KGN cells, however, all these results were reversed by miR-206 inhibitor. In conclusion, our findings suggested that liquiritin exerted anti-proliferative and apoptosis-inducing roles in KGN cells via miR-206/PI3K/AKT pathway, suggesting that liquiritin may be an effective therapeutic target for PCOS treatment.
RESUMEN
Endometriosis is an estrogen-dependent chronic gynecological syndrome. Recent studies have shown that long non-coding RNAs participate in the pathogenesis and development of endometriosis. This study aimed to explore the mechanisms of DHRS4 antisense RNA 1 (DHRS4-AS1) in endometriosis. Dual-luciferase reporter assays were conducted to determine the relationship between DHRS4-AS1, microRNA (miR)-139-5p, and arrestin domain-containing 3 (ARRDC3). Furthermore, the expression of DHRS4-AS1 and miR-139-5p in ectopic endometrial stromal cells (EC-ESCs) and endometriosis tissues was examined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Additionally, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), flow cytometry, and Transwell assays were performed to evaluate the proliferation, apoptosis, and migration and invasion of EC-ESCs, respectively. Western blotting and RT-qPCR were further utilized to determine cleaved-Caspase 3, Caspase 3, and matrix metalloproteinase 9 (MMP-9) expression levels. Compared with the EN group, DHRS4-AS1 levels were lower and miR-139-5p levels were higher in EC-ESCs and tissues obtained from patients with endometriosis. Functional assays validated that DHRS4-AS1 targets miR-139-5p, with ARRDC3 being a downstream target of miR-139-5p. Rescue experiments demonstrated that DHRS4-AS1 inhibited EC-ESC proliferation, migration, and invasion, but promoted apoptosis, by targeting miR-139-5p in endometriosis. cleaved-Caspase3 expression level and the cleaved-Caspase 3/Caspase 3 ratio increased, while the expression levels of MMP-9 decreased, after transfection with DHRS4-AS1 overexpression plasmids; however, the effects induced by DHRS4-AS1 overexpression could be partially reversed by co-transfection with the miR-139-5p mimic. The current study demonstrates that the DHRS4-AS1/miR-139-5p/ARRDC3 axis participates in the regulation of EC-ESC function.