长链非编码RNA GAS5、微小RNA-21在多囊卵巢综合征中的表达及生物学意义

Expressions of LncRNA GAS5 and miR-21 in polycystic ovary syndrome and their biological significance

目的探讨长链非编码RNA(LncRNA)GAS5与微小RNA-21(miR-21)在多囊卵巢综合征(PCOS)中的表达及其对卵巢颗粒细胞增殖及凋亡的影响。方法收集2017年4月至2018年10月于南阳市中心医院就诊的50例PCOS病人为观察组,选取同时期于南阳市中心医院就诊的45例非PCOS病人为对照组,实时荧光定量聚合酶链式反应(qRT-PCR)检测对照组与观察组病人血清中GAS5与miR-21的表达水平。采用Pearson法分析PCOS病人中GAS5与miR-21表达的相关性。原代培养大鼠卵巢颗粒细胞,分别将pcDNA、pcDNA-GAS5转染至卵泡颗粒细胞。CCK-8检测细胞增殖;流式细胞术检测细胞凋亡率;双荧光素酶报告实验验证GAS5与miR-21的靶向关系。结果观察组病人血清中GAS5的表达水平明显低于对照组[(0.52±0.11)比(1.00±0.12),P<0.05],miR-21的表达水平明显高于对照组[(2.36±0.57)比(1.01±0.10),P<0.05],GAS5与miR-21呈负相关(r=-0.310,P=0.028);TT、IL-18与RGAS5呈负相关(P<0.05),而与miR-21呈正相关(P<0.05);与pcDNA组相比,pcDNA-GAS5组细胞活力降低(P<0.05),细胞凋亡率升高[(36.49±5.21)比(8.52±1.20),P<0.05];双荧光素酶报告实验证实GAS5可靶向结合miR-21,并可负向调控miR-21的表达。结论GAS5过表达可能通过靶向调控miR-21表达从而抑制卵巢颗粒细胞增殖,促进细胞凋亡。

Objective To investigate the expression of LncRNA GAS5 and miR-21 in PCOS and their effects on the proliferation and apoptosis of ovarian granulosa cells.Methods qRT-PCR was used to detect the expression levels of GAS5 and miR-21 in the serum of patients in the control and observation groups.The correlation between GAS5 and miR-21 expression in PCOS patients was analyzed by Pearson method.Primary cultured rat ovarian granulosa cells were transfected with pcDNA,pcDNA-GAS5 into follicular granule cells,respectively.CCK-8 was used to detect cell proliferation.Apoptosis rate was measured by flow cytometry.The double luciferase report experiment verified the targeting relationship between GAS5 and miR-21.Results The expression level of GAS5 in the serum of the observation group was significantly lower than that of the control group[(0.52±0.11)vs.(1.00±0.12),P<0.05],and the expression level of miR-21 was significantly higher than that of the control group[(2.36±0.57)vs.(1.01±0.10),P<0.05].GAS5 was negatively correlated with miR-21(r=-0.310,P=0.028).GAS5 was negatively correlated with TT,IL-18(P<0.05),and miR-21 was positively correlated with TT,IL-18(P<0.05).Compared with the pcDNA group,the cell viability of the pcDNA-GAS5 group was significantly reduced(P<0.05),the apoptosis rate was significantly increased[(36.49±5.21)vs.(8.52±1.20),P<0.05].The double luciferase reporting experiment confirmed that GAS5 can target miR-21 and negatively regulate miR-21 expression.Conclusion GAS5 overexpression may inhibit miR-21 expression by targeting to inhibit ovarian granulosa cell proliferation and promote apoptosis.