DAPT抑制RM-1细胞增殖的实验研究 |
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引用本文: | 吴涛华,王春,林妙春,林永绥,刘成招,刘清平,张春.DAPT抑制RM-1细胞增殖的实验研究[J].中国现代医生,2012,50(11):10-12. |
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作者姓名: | 吴涛华 王春 林妙春 林永绥 刘成招 刘清平 张春 |
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作者单位: | 1. 福建医科大学附属闽东医院骨科,福建福安,355000 2. 福建医科大学附属闽东医院中心实验室,福建福安,355000 3. 福建医科大学附属闽东医院泌尿外科,福建福安,355000 |
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基金项目: | 福建省宁德市2011年科技计划项目(20110130) |
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摘 要: | 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。
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关 键 词: | γ-分泌酶抑制剂 Notch通路 前列腺癌细胞 增殖 细胞周期 |
The experimental study on suppressing proliferation of RM-1 cells by DAPT |
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Authors: | WU Taohua WANG Chun LIN Miaochun LIN Yongsui LIU Chengzhao LIU Qingping ZHANG Chun |
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Institution: | 1.Department of Orthopedics,Mindong Hospital Affiliated to Fujian Medical University,Fuan 355000,China;2.Department of Central Laboratory,Mindong Hospital Affiliated to Fujian Medical University,Fuan 355000,China;3.Department of Urological Surgery,Mindong Hospital Affiliated to Fujian Medical University,Fuan 355000,China |
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Abstract: | 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. |
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Keywords: | γ-Secretase inhibitor Notch signaling pathway RM-1 Cell proliferation Cell cycle |
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