摘要
目的观察雷公藤内酯醇对多发性骨髓瘤细胞RPMI 8226增殖和周期的影响,探讨细胞周期调控蛋白P21wap1/cip1和P27kip1在其中的作用和意义。方法采用MTT比色法和流式细胞术检测雷公藤内酯醇对RPMI 8226细胞增殖、凋亡和细胞周期的影响,半定量RT-PCR和Western blotting法检测RPMI 8226细胞中P21wap1/cip1、P27kip1 mRNA和蛋白表达。结果 雷公藤内酯醇能明显抑制RPMI 8226细胞增殖,其抑制作用呈时间、剂量依赖性,雷公藤内酯醇作用48 h的IC50值为(71.18±2.01)nmol/L。雷公藤内酯醇还可以诱导RP-MI 8226细胞周期阻滞于G0/G1期,随着雷公藤内酯醇浓度的增加,G0/G1期细胞逐渐增多,S期细胞逐渐减少。经雷公藤内酯醇干预后周期调节蛋白P21wap1/cip1和P27kip1的mRNA和蛋白表达水平明显上调。结论雷公藤内酯醇可以抑制RPMI 8226细胞增殖,该抑制作用是通过调控P21wap1/cip1和P27kip1的表达,从而阻止细胞周期G0/G1期过渡实现的。
Objective To investigate the effect of triptolide on cell proliferation,cell cycle distribution,DNA and protein expression regulation of P21wap1/cip1 and P27kip1 in human multiple myeloma RPMI 8226 cells.Methods Cell viability was detected by MTT assay and cell cycle distribution was measured by flow cytometry.Effect on mRNA expression of P21wap1/cip1 and P27kip1 was detected by RT-PCR.The change of protein expression was measured by Western blotting.Results Triptolide of varying concentration significantly induced the inhibition of proliferation in a dose-dependent manner and G0/G1 phase arrest of the cell cycle progression.The events were coincided with the upregulation of the mRNA and protein expression of P21wap1/cip1 and P27kip1.Conclusion These results suggest that triptolide might exhibit its strong antitumor effect via alternation of P21wap1/cip1 and P27kip1.It provides framework for a clinical evaluation of triptolide.
出处
《中草药》
CAS
CSCD
北大核心
2010年第11期1819-1823,共5页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(30700882)
