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长链非编码RNA A-30-P01019163对卵巢颗粒细胞βB2晶体蛋白表达的调控及对颗粒细胞增殖和凋亡的影响

LncRNA A-30-P01019163 regulates βB2 crystallin gene expression in ovarian granulosa cells and its effects on cell proliferation and apoptosis

    摘要
  • 摘要:
    目的 探讨lncRNA A-30-P01019163对卵巢颗粒细胞βB2晶体蛋白（βB2 crystallin，CryBB2）表达的调控作用，以及其对卵巢颗粒细胞增殖、凋亡的影响。
    方法 构建小鼠卵巢颗粒细胞过表达lncRNA A-30-P01019163体内模型，同时基于人卵巢颗粒细胞系KGN构建过表达或干扰lncRNA A-30-P01019163稳定细胞株。采用qRT-PCR和Western印迹法检测lncRNA A-30-P01019163过表达或干扰后颗粒细胞中CryBB2的表达水平，采用EdU法检测细胞增殖，采用TUNEL法和流式细胞术检测细胞凋亡。
    结果 随着小鼠周龄增长，卵巢颗粒细胞lncRNA A-30-P01019163表达逐渐增加（P < 0.05）。过表达lncRNA A-30-P01019163后，小鼠卵巢颗粒细胞中CryBB2 mRNA和蛋白表达均增加（P < 0.01）。对卵巢颗粒细胞系KGN过表达lncRNA A-30-P01019163后，其CryBB2 mRNA和蛋白表达均增加（P < 0.05）；颗粒细胞增殖加快，细胞周期相关蛋白D1及增殖细胞核抗原蛋白表达均增加（P < 0.01）；颗粒细胞凋亡增加，凋亡相关蛋白半胱氨酸蛋白酶3（caspase-3）切割体及多聚ADP-核糖聚合酶（poly ADP-ribose polymerase，PARP）切割体表达均增加（P < 0.05）。而干扰卵巢颗粒细胞系KGN中lncRNA A-30-P01019163后，CryBB2 mRNA和蛋白表达均下调（P < 0.01）；颗粒细胞增殖减慢，细胞周期相关蛋白D1及增殖细胞核抗原蛋白表达均降低（P < 0.01）；颗粒细胞凋亡减少，caspase-3及PARP切割体表达均降低（P < 0.05）。
    结论 小鼠卵巢颗粒细胞中lncRNA A-30-P01019163表达随卵巢发育逐渐增加，参与调控CryBB2表达，并与颗粒细胞增殖、凋亡密切相关。

     

    Abstract:
    Objective To explore the regulatory effect of lncRNA A-30-P01019163 on the expression of ovarian granulocytes βB2 crystallin (CryBB2) and its effects on ovarian granulocyte proliferation and apoptosis.
    Methods The model of over-expression lncRNA A-30-P01019163 was constructed, and a stable cell line based on human ovarian granulosa cell line KGN with of overexpression or interference lncRNA A-30-P01019163 was established. The expression level of CryBB2 in after expression or interference was detected by qRT-PCR and protein blotting method, and the cell proliferation was detected by EdU, and apoptosis was detected by TUNEL and flow cytometric analysis.
    Results As the mice age increasesd, the expression of lncRNA A-30-P01019163 in the granulocytes gradually increasesd (P < 0.05). After overexpression of lncRNA A-30-P01019163 in KGN cells, the CryBB2 mRNA and protein expressions increased (P < 0.05); granulocyte proliferation accelerated, cell cycle-related protein D1 and proliferating cell nuclear antigen expressions increased (P < 0.01); the apoptosis of granulocytes increased, and cleaved caspase-3 and poly ADP-ribose polymerase (PARP) expressions were all increased(P < 0.05). After interfering lncRNA A-30-P01019163 in KGN cells, CryBB2 mRNA and protein expressions were reduced (P < 0.01); cell proliferation slowed down, cell cycle-related protein D1 and proliferating cell nuclear antigen expressions decreased (P < 0.01); cell apoptosis decreased, cleaved caspase-3 and PARP expressions decreased (P < 0.05).
    Conclusions The expression of lncRNA A-30-P01019163 increased along with ovarian granulosa cell development and is involved in the regulation of CryBB2 protein expression. LncRNA A-30-P01019163 is closely related to proliferation and apoptosis of ovarian granulosa cells.

     

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