摘要
目的采用定量和定性相结合的方法观察丹皮酚对HT-29细胞的增殖抑制作用并对其可能的作用机制进行探讨。方法用MTF法和流式细胞仪定量测定丹皮酚对HT-29细胞的抑制率及细胞凋亡率，选用TUNEL法及细胞免疫组化从形态学方面探寻丹皮酚对HT-29细胞作用的可能机制。结果丹皮酚在0．024～1．504mol·L^-1剂量下对体外培养的大肠癌HT-29细胞的增殖均有抑制作用，呈现明显的浓度效应关系；在此浓度（0．024～1．504mol·L^-1），分别作用24，48，72和96h，抑制率随之增加，呈明显的时间效应关系；经1．504mol·L^-1的丹皮酚处理的HT-29细胞，细胞凋亡率明显增加与阴性对照有明显差异；流式细胞仪检测在G．期前出现sub—G1峰，S期细胞增多，G1、G2期细胞减少，随着丹皮酚浓度的增加，HT-29细胞的凋亡率逐渐增加，每个实验组的凋亡率与对照组相比差异有显著性（P〈0．05）；丹皮酚作用HT-29细胞可使Fas／FasL表达增加。结论丹皮酚可能通过七调大肠癌HT-29细胞Fas／FasL蛋白的表达而诱导细胞凋亡是其抑制大肠癌细胞增殖的机制之一．
Aim To investigate the effects of paeonol （Pae） in inhibiting the proliferation of HT-29 cell and probe into the possible molecule mechanism by quantitative and qualitative assay. Methods The inhibited rate and apoptotic rate of HT-29 cells were measured quantitatively by MTT assay. We were trying to find out the possible mechanism through the morphologic observation on paeonol-processed HT-29 cell line by TUNEL assay and immunocytochenical method. Resuits Pae, in the concentration of 0. 024 - 1. 504μmol·L^-1, inhibited the proliferation of HT-29 cells in vitro, which showed obvious concentration-effect relationship;The inhibited rate of HT-29 cells was also increased when Pae （0. 024 - 1. 504μmol·L^-1） was treated for 24,48,72 and 96 h,which showed obvious time-effect relationship. After treated with the concentration of 1. 504μmol·L^-1, the apoptotic rate of HT-29 cells significantly increased, which showed significant difference compared with control; We examined all the experimental groups by flow cytometry, which showed that sub-G1 peak appeared before G1 period and the cells in S period increased, while the cells in G1, G2 period decreased, the apoptotic rate of HT-29 cells gradually increased along with the increasing of paeonol concentration. The apoptotic rate in experimental groups vs control has significant difference（ P 〈 0. 05 ） ; Pae up-regulated the expression of Fas/FasL protein in colorectal cancer cells. Conclusion Pae can inhibit the proliferation of the HT-29 cells by up-regulating the expression of Fas/FasL protein in colorectal cancer cells .
出处
《中国药理学通报》
CAS
CSCD
北大核心
2005年第10期1251-1254,共4页
Chinese Pharmacological Bulletin
基金
湖北省自然基金资助项目(No2004ABA243)
