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孕激素诱导的microRNA-152对子宫内膜上皮细胞增殖的抑制作用

Effect of Progesterone-induced MicroRNA-152 on the Proliferation of Endometrial Epithelial Cells

  • 摘要: 目的 研究孕激素诱导microRNA-152的表达及其对子宫内膜上皮细胞增殖的抑制作用。方法 培养子宫内膜上皮细胞Ishikawa,将其分为4组,分别为空白对照组(control,C组)、10-8mol/L雌激素处理组(estrogen,E组)、10-6mol/L孕激素处理组(progesterone,P组)以及相应浓度的雌、孕激素联合处理组(estrogen plus progesterone,E&P组),利用实时定量PCR检测microRNA-152的成熟体microRNA-152-3p的表达。将microRNA-152-3p拟似剂转染雌激素预处理的细胞,microRNA-152-3p抑制剂转染雌、孕激素联合处理的细胞,CCK-8法观察microRNA-152-3p对细胞增殖的影响。利用microRNA靶蛋白数据库预测microRNA-152-3p的靶蛋白,并利用Western blot加以验证。结果 实时定量PCR结果显示E组与C组相比,其细胞microRNA-152-3p的表达量无明显变化( P>0.05);P 组细胞microRNA-152-3p的表达量相比C组增加( P<0.05);且E&P组的microRNA-152-3p表达量高于C组和P组( P均<0.05)。对microRNA-152-3p靶蛋白的预测表明CDC14A是其一个重要的靶蛋白。转染microRNA-152-3p拟似剂能够抑制雌激素对子宫内膜上皮细胞的增殖效应( P<0.05),同时下调CDC14A蛋白的表达( P<0.05)。而microRNA-152-3p抑制剂能一定程度解除孕激素对子宫内膜上皮细胞的增殖抑制效应( P<0.05),同时上调CDC14A蛋白水平( P<0.05)。结论 孕激素可通过诱导microRNA-152-3p的表达抑制子宫内膜上皮细胞的增殖,CDC14A蛋白可能是microRNA-152-3p抑制子宫内膜上皮细胞增殖的靶蛋白。

     

    Abstract: Objective To determine the expression of microRNA-152 induced by progesterone and its effect on the proliferation of endometrial epithelial cells (EECs). Methods Cultured EECs, Ishikawa were divided into four groups: control group (C group), 10-8mol/L estrogen treated group (E group), 10-6mol/L progesterone group (P group) and estrogen plus progesterone treated group (E&P group). The expression of mature microRNA-152 (microRNA-152-3p)of was detected by qRT-PCR. The estrogen treated cells were transfected with mimic-microRNA-152-3p. The estrogen and progesterone treated cells were transfected with inhibitor-microRNA-152-3p. Cell proliferations were detected by CCK-8 assay. The target gene of microRNA-152-3p proteins was predicted using microRNA target databases and validated by Western blot. Results qRT-PCR showed no difference between C and E groups ( P>0.05) in the expression of microRNA-152-3p. P group had higher expressions of microRNA-152-3p than C group ( P<0.05). E&P group had higher expressions of microRNA-152-3p than C group and P group. MicroRNA target protein prediction suggested that CDC14A is one of direct target proteins of microRNA-152-3p. The results of CCK-8 assay showed that mimic-microRNA-152-3p transfection blocked proliferations of estrogen treated cells and lowered expressions of CDC14A in these cells; while inhibitor-microRNA-152-3p promotes proliferations of estrogen and progesterone treated cells and increased expressions of CDC14A in these cells. Conclusion Progesterone may suppress proliferations of EECs through inducing expressions of microRNA-152-3p. CDC14A is probably one target protein of microRNA-152-3p for its action on EECs.

     

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