Aberrant Expression Of LINC00477/miR-128/SOCS6 Axis And Related Mechanism In Polycystic Ovary Syndrome

Background:Polycystic ovary syndrome(PCOS)is a heterogeneous endocrine disease frequently diagnosed among women,affecting 5%-10%of women at childbearing period.But the detailed pathogenesis is unclear.Diverse researches have demonstrated the function of long non-coding RNAs(lncRNAs)in the modulation of PCOS development,and explored the associated molecule and pathway regulation mechanism.Reports have confirmed that LINC00477 functioned in gastric cancer.It has been reported that LINC00477 was over-expressed in PCOS,however,the specific mechanism remains unclear.Objective:To probe the expression pattern of LINC00477 in the serum of PCOS patients,to verify the elevated expression of LINC00477 in PCOS animal models,and to explore the impact of LINC00477 change on proliferation and apoptosis capacities of human granulosa cells.Eventually,the mechanism that LINC00477 involved in was investigated.Methods:Serum samples of PCOS patients and healthy controls at our hospital from January,2019 to April,2019 were collected.Quantitative real-time PCR(qRT-PCR)estimated the level of LINC00477,and its relationship with the prognosis of patients was also analyzed.In situ hybridization(ISH)and qRT-PCR dissected LINC00477 expression in ovarium or serum of PCOS and normal mice.Granulosa cells were transfected and divided into four groups:blank control(vector)group,LINC00477 overexpression(LINC00477 overexpression vector)group,negative control group(si-NC),and LINC00477 knockdown(si-LINC00477 1/2#)group.MTT colorimetric assay,colony formation assay,as well as flow cytometry separately measured the impact of LINC00477 expression change on cell viability,proliferation and apoptosis.The level of miR-128 in serum of PCOS patients and healthy controls was explored via qRT-PCR,and its correlation with LINC00477 was further dissected.qRT-PCR also examined miR-128 expression in ovarium or serum of PCOS and normal mice.Bioinformatics predicted the binding between LINC00477 and miR-128,which was affirmed via dual luciferase reporter and RNA pull-down assays.In rescue experiments,cells were divided into blank control group,LINC00477 overexpression group,miR-NC(LINC00477 overexpression vector and miR-NC)group,and miR-128(LINC00477 overexpression vector and miR-128 mimics)group.The above function experiments were conducted to test cell proliferation and apoptosis.The level of SOCS6 in serum of PCOS patients and healthy controls was explored via qRT-PCR,and its correlation with miR-128 was further dissected.qRT-PCR also examined miR-128 expression in ovarium or serum of PCOS and normal mice.In rescue experiments,cells were divided into blank control group,miR-128 overexpression group,miR-NC group,and miR-128 mimics+SOCS6 group.The above function experiments were conducted to test cell proliferation and apoptosis.Results:LINC00477 was significantly heightened in serum of 90 PCOS patients.Both the ovarium and serum of PCOS mice models exhibited higher level of LINC00477 than those of normal mice.In granulosa cells,overexpression of LINC00477 hindered proliferation and boosted apoptosis capacities.In contrast,its silencing accelerated proliferation and decreased apoptosis.miR-128,target of LINC00477,was overtly decreased in PCOS patients.Besides,distinctly reduced miR-128 expression was observed in the ovarium and serum of PCOS mice models.Mechanism assays validated the binding between LINC00477 and miR-128.In rescue experiments,elevation of miR-128 eliminated the impacts of LINC00477 overexpression on proliferation and apoptosis of granulosa cells.SOCS6,target of miR-128,was overtly increased in PCOS patients.Besides,overexpression of SOCS6 was observed in the ovarium and serum of PCOS mice models.Mechanism assays validated the binding between SOCS6 and miR-128.In rescue experiments,elevation of SOCS6 eliminated the impacts of miR128 overexpression on proliferation and apoptosis of granulosa cells.Conclusion:1.The expression of LINC00477 was significantly up-regulated in serum of PCOS patients,ovarian tissue and serum of PCOS mouse model;Inhibit the activity and proliferation of ovarian granulosa cells and promote apoptosis.2.The expression of miR-128 in serum of PCOS patients,ovarian tissue and serum of PCOS mouse model was significantly down-regulated,which was negatively correlated with the expression of LINC00477.LINC00477 in ovarian granulosa cells affects cell viability,proliferation and apoptosis by negatively regulating miR-128.3.The expression of socs6 was significantly up-regulated in serum of PCOS patients,ovarian tissue and serum of PCOS mouse model,which was negatively correlated with the expression of miR-128.MiR-128 in ovarian granulosa cells negatively regulates the activity,proliferation and apoptosis of socs6 cells.4.LINC00477/miR-128/SOCS6 is involved in the regulation of proliferation and apoptosis of ovarian granulosa cells and plays an important role in the occurrence of PCOS.