沉默环状RNA_单酰甘油酯酶促进睾丸支持细胞增殖而抑制凋亡

Silencing circular RNA_monoglyceride lipase promotes the proliferation and inhibits apoptosis of sertoli cell in tesis

目的探讨沉默环状RNA_单酰甘油酯酶(Circular RNA_monoglyceride lipase,circ_MGLL)表达对睾丸支持细胞增殖和凋亡的影响及潜在机制。方法应用siRNA沉默睾丸支持细胞中circ_MGLL和MGLL的表达。应用CCK-8和EdU检测沉默circ_MGLL对支持细胞增殖能力的影响。应用流式细胞术检测沉默circ_MGLL对支持细胞周期和凋亡的影响。应用生信网站预测与circ_MGLL具有结合潜能的miRNA。应用荧光素酶报告实验检测circ_MGLL与相关miRNAs的结合能力。结果沉默circ_MGLL的表达后,支持细胞增殖速度加快、增殖细胞比例和S期细胞比例升高(均P<0.05),而凋亡细胞比例和G1期细胞比例降低(P<0.05)。机制方面研究显示,circ_MGLL可结合miR-1228、miR-1233、miR-149和miR-924。结论circ_MGLL可结合miR-1228/miR-1233/miR-149/miR-924,并促进睾丸支持细胞增殖而抑制细胞凋亡。

Objective To explore the effects and underlying mechanism of silencing circ_MGLL on the proliferation and apoptosis in sertoli cells.Methods Small interfering RNA(siRNA)was used to silence the expression of circ_MGLL in sertoli cells.CCK-8 and EdU were used to detect the effects of circ_MGLL on the proliferation in sertoli cells.Flow cytometry was used to detect the effects of circ_MGLL on the apoptosis and cell cycle distribution in sertoli cells.Bioinformatics website was used to predict the potential binding miRNAs of circ_MGLL.Luciferase reporter assays was used to detect the binding ability between circ_MGLL and miRNAs.Results After silencing the expression of circ_MGLL,the proliferating speed of sertoli cells were increased,the proportion of proliferating cells and cells at S stage were up-regulated,while the levels of apoptotic cells were decreased and cells at G1 stage were down-regulated.Mechanism study found circ_MGLL could bind with miR-1228,miR-1233、miR-149 and miR-924.Conclusion circ_MGLL binds with miR-1228/miR-1233/miR-149/miR-924,and inhibits cell proliferation and induces cell apoptosis.