lnc81在卵巢颗粒细胞中的生物学作用及机制研究

Study on the biological function and mechanism of lnc81 in ovarian granulosa cells

[目的]探讨lnc81在卵巢颗粒细胞中的生物学作用及机制。[方法]收集于医院接受体外受精-胚胎移植治疗78例不孕患者卵泡液,分离人原代卵巢颗粒细胞。构建lnc81干扰和过表达模型,分为对照组、干扰组和过表达组。分析lnc81表达对人卵巢颗粒细胞增殖、凋亡及Hippo信号通路蛋白表达的影响。[结果]lnc81明确表达于人原代卵巢颗粒细胞中,且主要表达于细胞核。过表达组细胞增殖水平(0.77±0.09)明显高于干扰组(0.41±0.08)和对照组(0.63±0.12),差异有统计学意义(P<0.05)。干扰组细胞凋亡率(5.93±0.18)高于对照组(2.36±0.23)和过表达组(1.14±0.21),差异有统计学意义(P<0.05)。干扰组细胞BAX/BCL-2蛋白水平比率(0.97±0.06)高于对照组(0.52±0.04)以及过表达组(0.36±0.05),差异有统计学意义(P<0.05)。干扰组细胞CCN1、CCN2蛋白水平[(8.64±1.23),(5.69±0.82)]高于对照组[(5.36±1.14),(4.12±0.78)]和过表达组[(3.23±1.08),(3.08±0.76)],差异有统计学意义(P<0.05)。[结论]lnc81通过上调CCN1、CCN2蛋白表达抑制卵巢颗粒细胞增殖,加速其凋亡。

[Objective]To investigate the biological effects and mechanisms of long non-coding RNA 81(lnc81)in ovarian granulosa cells.[Method]Follicular fluid was collected from 78 infertile patients undergoing in vitro fertilization and embryo transfer(IVF-ET).Lnc81 interference and overexpression models were constructed,which were divided into control group,interference group and overexpression group.The effects of lnc81 expression on proliferation,apoptosis and expression of Hippo signaling pathway in human ovarian granulosa cells were analyzed.[Result]Lnc81 was expressed in human primary ovarian granulosa cells and mainly in the nucleus.The cell proliferation level in the overexpression group was(0.770±09)was significantly higher than the interference group(0.410±08)and control group 0.630±The difference was statistically significant(P<0.05).In the interference group,the apoptosis rate was(5.931±18)was higher than that of the control group(2.360±23)and overexpression group(1.140±21),the difference was statistically significant(P<0.05).The ratio of Bax/Bcl-2 protein level in the interference group was(0.970±06)was higher than that of the control group(0.520±04)and overexpression group(0.360±The difference was statistically significant(P<0.05).CCN1 and CCN2 protein levels in the interference group[(8.641±1.23),(5.69±82)]was higher than that in the control group[(5.360±1.14),(4.12±78)]and overexpression group[(3.230±1.08),(3.08±76)],the difference was statistically significant(P<0.05).[Conclusion]Lnc81 inhibits the proliferation and accelerates the apoptosis of ovarian granulosa cells by up regulating the expression of CCN1 and CCN2.