Long non-coding RNA HLA-F antisense RNA 1 inhibits the maturation of microRNA-613 in polycystic ovary syndrome to promote ovarian granulosa cell proliferation and inhibit cell apoptosis

MicroRNA-613 (miR-613) inhibits granulosa cell proliferation, suggesting its involvement in polycystic ovary syndrome (PCOS). We predicted that long non-coding RNA (lncRNA) HLA-F antisense RNA 1 (HLA-F-AS1) could interact with premature miR-613. We then explored the crosstalk between HLA-F-AS1 and miR-613 in PCOS. In this study, follicular fluid donated by 58 healthy controls and 58 PCOS patients was used to analyze the expression of HLA-F-AS1 and miR-613 (mature and premature). The direct interaction between HLA-F-AS1 and premature miR-613 was evaluated by RNA pull-down assay. Overexpression of both HLA-F-AS1 and miR-613 was achieved in granulosa cells to assess their interactions. Cell proliferation and apoptosis were detected with BrdU assay and cell apoptosis assay, respectively. We found that miR-613 was highly expressed in PCOS, while HLA-F-AS1 was downregulated in PCOS. HLA-F-AS1 directly interacted with premature miR-613, and overexpression of HLA-F-AS1 increased the expression levels of premature miR-613, but decreased the expression levels of mature miR-613. HLA-F-AS1 increased ovarian granulosa cell proliferation and inhibited cell apoptosis. MiR-613 played an opposite role and suppressed the role of HLA-F-AS1. Therefore, HLA-F-AS1 may inhibit the maturation of miR-613 in PCOS to promote ovarian granulosa cell proliferation and inhibit cell apoptosis.

微小RNA-613（MicroRNA-613，miR-613）可抑制颗粒细胞增殖，提示其参与多囊卵巢综合征（polycystic ovary syndrome，PCOS）的病理过程。本研究预测长链非编码RNA（long non-coding RNA，lncRNA）HLA-F反义RNA1（HLA-F antisense RNA 1，HLA-F-AS1）可与前体miR-613相互作用，随后探讨了PCOS中HLA-F-AS1与miR-613之间的串扰调控关系。本研究纳入58名健康对照者与58名PCOS患者的卵泡液，用于检测HLA-F-AS1以及miR-613（成熟体与前体）的表达水平。通过RNA下拉实验（RNA pull-down assay）评估HLA-F-AS1与前体miR-613之间的直接相互作用；在颗粒细胞中分别过表达HLA-F-AS1与miR-613，以验证二者的相互调控作用。分别采用BrdU实验（BrdU assay）与细胞凋亡实验（cell apoptosis assay）检测细胞增殖与凋亡情况。结果显示，miR-613在PCOS患者样本中呈高表达，而HLA-F-AS1在PCOS患者样本中表达下调。HLA-F-AS1可与前体miR-613直接结合；过表达HLA-F-AS1可提升前体miR-613的表达水平，却降低成熟miR-613的表达水平。HLA-F-AS1可促进卵巢颗粒细胞增殖并抑制细胞凋亡，而miR-613则发挥相反的生物学功能，且可抑制HLA-F-AS1的上述作用。综上，HLA-F-AS1或可通过抑制PCOS中miR-613的成熟过程，从而促进卵巢颗粒细胞增殖并抑制细胞凋亡。