DAPT抑制RM-1细胞增殖的实验研究

目的研究Notch信号通路特异性阻断剂DAPT对小鼠前列腺癌RM-1细胞增殖的影响。方法体外培养RM-1细胞,实验组以不同浓度DAPT（0.25、0.5、1、2、5μmol/L）处理,对照组不加DAPT;利用MTT法检测细胞增殖,流式细胞术检测细胞周期（PI法）,RT-PCR法检测Notch1、Jagged1基因表达。结果与对照组比较,0.25μmol/L剂量组对RM-1细胞增殖、周期、Notch1及Jagged1 mRNA表达的影响无差异;0.5、1、2、5μmol/L剂量组均能显著抑制RM-1细胞增殖,随着DAPT浓度的增加,G0/G1期细胞比例逐渐增多,S期细胞比例逐渐减少,G2/M期细胞比例无明显差异,Notch1、Jagged1 mRNA的表达逐渐下调。结论当DAPT浓度≥0.5μmol/L时能够抑制RM-1细胞的增殖,且呈剂量依赖性,其机制可能与诱导细胞G0/G1期阻滞、Notch1及Jagged1 mRNA的表达下调有关。

The experimental study on suppressing proliferation of RM-1 cells by DAPT

Objective To investigate the effect of DAPT(γ-secretase inhibitor) on the proliferation of mice prostate cancer RM-1 cells by inhibiting Notch signaling pathway in vitro.Methods RM-1 cells were randomly divided into two groups: the experimental group in which specimens were treated with DAPT at the concentration of 0.25,0.5,1,2,5 μmol/L,and the control group in which specimens were treated with medium;Cell proliferation was determined using MTT method.The cell cycle was analyzed by flow cytometry(FCM) with PI staining method;RT-PCR was used to confirm the expression of Notch1/Jagged1 mRNA.Results Compared with the control group,0.25 μmol/L dose group had no significant effects on proliferation,cell cycle distribution and the expressions of Notch1/Jagged1 mRNA;0.5,1,2,5 μmol/L dose group significantly inhibited RM-1 cell proliferation,and with the increase of the concentration,G0/G1 cell cycle increased gradually,S cell cycle gradually reduced,G2/M cell cycle had no obvious difference,and the expression of Notch1/Jagged1 mRNA down-regulated gradually.Conclusion When the concentration of DAPT ≥0.5 μmol/L,DAPT can significantly inhibit the proliferation of RM-1 cells in dose-dependent manner in vitro,which is mechanistically linked with inducing G0/G1 cycle arrest and down-regulation of Notch1/Jagged1.