瑞芬太尼通过上调lncRNA DGCR5表达影响卵巢癌细胞增殖和转移的机制研究

Mechanism of remifentanil affects proliferation and metastasis of ovarian cancer cells by up-regulating expression of lncRNA DGCR5

  • 摘要:
    目的 探讨瑞芬太尼对长链非编码RNA(lncRNA)DiGeorge综合征关键区域基因5(DGCR5)表达的调控及卵巢癌细胞生长和转移的影响。
    方法 采用细胞计数试剂盒8 (CCK-8)法检测0、0.5、5.0、50.0、500.0 ng/mL瑞芬太尼组、si-NC组、si-lncRNA DGCR5组、si-NC+500 ng/mL瑞芬太尼组、si-lncRNA DGCR5+500 ng/mL瑞芬太尼组卵巢癌细胞SKOV3、OVCAR3的增殖; 采用流式细胞仪、Transwell法、免疫印迹试验(Western blot)和定量逆转录聚合酶链反应(qRT-PCR)依次检测SKOV3、OVCAR3细胞的凋亡、迁移、侵袭以及细胞周期蛋白D1(CyclinD1)、活化的天冬氨酸特异性半胱氨酸蛋白酶-3(Cleaved-caspase-3)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)的表达和lncRNA DGCR5表达。
    结果 与0 ng/mL瑞芬太尼比较, 0.5、5.0、50.0、500.0 ng/mL瑞芬太尼可降低SKOV3、OVCAR3细胞的增殖活性、迁移及侵袭细胞数量以及CyclinD1、MMP2、MMP9蛋白表达水平, 增加凋亡率、Cleaved-caspase-3蛋白和lncRNA DGCR5表达水平,差异有统计学意义(P < 0.01)。与si-NC组比较, lncRNA DGCR5低表达可使卵巢癌细胞SKOV3和OVCAR3的lncRNA DGCR5表达水平、凋亡率、Cleaved-caspase-3蛋白表达水平降低,细胞增殖活性、迁移及侵袭数量、CyclinD1、MMP2、MMP9蛋白表达水平升高,差异有统计学意义(P < 0.01)。lncRNA DGCR5低表达可以逆转瑞芬太尼对卵巢癌细胞SKOV3和OVCAR3增殖、迁移、侵袭、凋亡的影响。
    结论 瑞芬太尼通过上调lncRNA DGCR5, 抑制卵巢癌细胞SKOV3、OVCAR3的增殖和转移,促进其凋亡。

     

    Abstract:
    Objective To explore the regulation of remifentanil on the expression of long non-coding RNA (lncRNA) DiGeorge syndrome critical region gene 5 (DGCR5) and its effect on growth and metastasis of ovarian cancer cells.
    Methods Cell Counting Kit 8 (CCK-8) method was used to detect the proliferation of ovarian cancer cells SKOV3 and OVCAR3 in the 0, 0.5, 5.0, 50.0 and 500.0 ng/mL remifentanil group, si-NC group, si-lncRNA DGCR5 group, si-NC+500 ng/mL remifentanil group, and si-lncRNA DGCR5+500 ng/mL remifentanil group; the flow cytometry, Transwell method, Western blot and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were sequentially used to determine cell apoptosis, cell migration, cell invasion, CyclinD1, activated aspartic acid specific cysteine protease-3 (Cleaved-caspase-3), matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9) expressions and lncRNA DGCR5 expression in SKOV3 and OVCAR3 cells.
    Results Compared with 0 ng/mL remifentanil, remifentanil at concentrations of 0.5, 5.0, 50.0 and 500.0 ng/mL was able to significantly reduce the proliferation activity, numbers of migration andinvasion in SKOV3 and OVCAR3 cells as well as the protein expression levels of CyclinD1, MMP2 and MMP9, and significantly increase apoptosis rate and expression levels of Cleave-caspase-3 protein and lncRNA DGCR5 (P < 0.01). Compared with the si-NC group, low expression of lncRNA DGCR5 was able to significantly decrease the expression level of lncRNA DGCR5, apoptosis rate and expression level of Cleaved-caspase-3 protein in ovarian cancer cells SKOV3 and OVCAR3, and significantly increase cell proliferation activity, numbers of migration and invasion, and the protein expression levels of CyclinD1, MMP2 and MMP9 (P < 0.01). The low expression of lncRNA DGCR5 was able to reverse the effect of remifentanil on the proliferation, migration, invasion and apoptosis of ovarian cancer cells SKOV3 and OVCAR3.
    Conclusion Remifentanil can inhibit the proliferation and metastasis of ovarian cancer cells SKOV3 and OVCAR3, and promote apoptosis of cells by up-regulating lncRNA DGCR5.

     

/

返回文章
返回