摘要
目的研究PPARγ激动剂罗格列酮(ROZ)与维A酸联合应用对人乳腺癌MCF-7细胞相关蛋白表达的影响,阐明PPARγ激活后的抗肿瘤作用以及相关机制。方法MTT法检测ROZ、维A酸及两者联合应用对MCF-7细胞生长的作用。用荧光染料Hochest33342对细胞单染观察MCF-7细胞凋亡形态。用流式细胞仪检测MCF-7细胞的细胞周期分布。采用Western Blot法测定药物对MCF-7细胞相关蛋白表达的影响。结果ROZ对MCF-7细胞有一定的生长抑制作用,与维A酸合用后能显著抑制MCF-7细胞增殖,且两者呈协同作用(q>1.15)。维A酸能增强ROZ的凋亡诱导作用,表现为核碎裂和核固缩的细胞增多。流式细胞术分析二者合用可导致G1期细胞周期阻滞,S期细胞减少。Western Blot结果显示,ROZ可促进PPARγ、P53、Erk1/2、p-ERK1/2及JNK的表达,抑制ERα、C-myc及p-JNK的表达;与维A酸联合应用时可显著升高PPARγ表达,降低ERα、Bcl-2、C-myc及p-JNK表达的作用增强,并且抑制ERK的磷酸化。结论ROZ与维A酸联合应用可显著抑制MCF-7细胞的增殖,具有协同作用,可通过对Bcl-2、C-myc、P53、ERα和MAPK信号传导通路的影响而发挥抑制细胞增殖、诱导凋亡的作用。
Abstract
OBJECTIVE To study the effect of combination of PPARγ agonist Rosiglitazone(ROZ) with tretinoin on the relative protein expression in MCF-7, to study their anti-cancer activity and the mechanism of action. METHODS MTT assay was used to examine the growth activities of MCF-7 cells. The cooperative effects of ROZ and tretinoin on inhibiting the growth of MCF -7 cells were analyzed. Morphological characteristic of apoptosis in MCF-7 cells was detected by stain with Hochest 33342.Flow cytometry(FCM) was used to examine the effects of ROZ alone and ROZ combined with tretinoin on changes in cell cycle.Western Blot was used for the detection of PPARγ,ERα,p53,Bcl-2,C-myc,ERK,p-ERK,JNK and p-JNK protein. RESULTS ROZ produced antiproliferative effect on MCF-7 cells. Enhanced antiproliferative effects were obtained when MCF-7 cells were treated with ROZ combined with tretinoin.The combination induced apoptosis and G1 cell cycle arrest.Morphologically,combination of ROZ with tretinoin dramatically induced typical apoptotic change-karyorrhexis in MCF-7 cells,whereas the effect of ROZ alone was barely.The data of Western Blot showed that ROZ increased PPARγ,p53,Erk1/2,p-ERK1/2 and JNK protein level,decreased ERα,C-myc and p-JNK protein level. Combination with ROZ and tretinoin caused PPARγ protein level to increase markedly and caused ERα,Bcl-2,C-myc,p-JNK and p-ERK to decrease significantly. There were no significant change for p53,ERK and JNK protein level.CONCLUSION The combination of ROZ and tretinoin in MCF-7 cells demonstrated a cooperative effect on growth inhibition and apoptosis induction. In addition, with PPARγ actived,the regulation of p53,Bcl-2,C-myc,ERK,JNK,p-JNK and p-ERK contributed to growth inhibition and apoptosis induction.
关键词
过氧化物酶体激活物增殖受体 /
罗格列酮 /
维A酸 /
MCF-7
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Key words
peroxisome proliferator-activated receptors /
rosiglitazone /
tretinoin /
MCF-7 /
the expression of protein
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季宇彬;蔡林;陈晓光.
罗格列酮与维A酸联合应用对MCF-7细胞相关蛋白表达的影响[J]. 中国药学杂志, 2008, 43(16): 1221-1227
JI Yu-in;CI Lin;CHEN Xio-gung.
Effect of Combination of PPARγ Agonist Rosiglitazone with Tretinoin on the Relative Protein Expression in MCF-7 Cells [J]. Chinese Pharmaceutical Journal, 2008, 43(16): 1221-1227
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脚注
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基金
国家自然科学基金资助项目(30400352)
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