瑞芬太尼抑制人肝癌细胞Notch信号通路及细胞生长的研究

Study on remifentanilin inhibiting Notch signal pathway and cell growth of human hepatocarcinoma cells

  • 摘要:
      目的  探讨瑞芬太尼对肝癌细胞增殖、周期和凋亡及其相关Notch信号通路的影响。
      方法  通过体外培养肝癌细胞HepG2, 采取不同浓度(0、5、50、500 ng/mL)的瑞芬太尼处理细胞,并设为瑞芬太尼各剂量组。采用500 ng/mL瑞芬太尼和20 μmol/L Notch通路抑制剂DAPT处理细胞,分别设为瑞芬太尼组、瑞芬太尼+DAPT组。采用CCK8检测细胞的增殖; 流式细胞术检测细胞周期和凋亡; Western blot检测p27、Cyclin D1、Bax、cleaved cas-3和Notch通路相关蛋白的表达。
      结果  0、5、50、500 ng/mL的瑞芬太尼均可以降低肝癌细胞的450 nm处的光密度值(OD450 nm值)、S期细胞比率,增加G0/G1期细胞比率、细胞凋亡率,上调p27、Bax、cleaved cas-3蛋白表达,下调Cyclin D1、Notch1、Hes1蛋白表达,并呈浓度依赖性。瑞芬太尼+DAPT组的OD450 nm值、S期细胞比率、Cyclin D1蛋白表达低于瑞芬太尼组, G0/G1期细胞比率、细胞凋亡率、p27、Bax、cleaved cas-3蛋白表达高于瑞芬太尼组,差异均有统计学意义(P < 0.05)。
      结论  瑞芬太尼抑制肝癌细胞增殖,阻滞细胞周期,并诱导细胞凋亡,且抑制Notch信号通路。

     

    Abstract:
      Objective  To explore the effects of remifentanil on the proliferation, cycle and apoptosis of hepatocellular carcinoma cells as well as the related Notch signaling pathway.
      Methods  Hepatocellular cancer cells HepG2 were cultured in vitro and treated with remifentanil at different concentrations (0, 5, 50, 500 ng/mL), which were recorded as the remifentanil dose groups. Cells were treated with 500 ng/mL remifentanil and 20 μmol/L Notch pathway inhibitor DAPT, which were named as remifentanil group and remifentanil+DAPT group respectively. CCK8 was used to detect cell proliferation; flow cytometry was used to detect cell cycle and apoptosis; western blot was used to detect the expressions of p27, Cyclin D1, Bax, cleaved cas-3, and Notch pathway related proteins.
      Results  Remifentanil at concentrations of 0, 5, 50 and 500 ng/mL could reduce the optical density value at 450 nm (OD450 nm value) and S stage cell ratio, increase the G0/G1 stage cell ratio and apoptosis rate, up regulate the expressions of p27, Bax and cleaved CAS-3 proteins, and downregulate the expressions of Cyclin D1, Notch1 and Hes1 proteins in a concentration dependent manner. The OD450 nm value, S stage cell ratio and expression of Cyclin D1 protein in remifentanil+DAPT group were significantly lower than those in remifentanil group, and the G0/G1 stage cell ratio, apoptosis rate, and expressions of p27, Bax and cleaved CAS-3 proteins were significantly higher than those in remifentanil group (P < 0.05).
      Conclusion  Remifentanil can inhibit the proliferation of hepatocarcinoma cells, block cell cycle, induce apoptosis and inhibit Notch signaling pathway.

     

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