Zhang Gui-Ping, Lu Yin-Yan, Lv Jia-Chun, Ou Hui-Jian
Guangzhou Medical College, Guangzhou 510182, China.
Zhongguo Zhong Yao Za Zhi. 2006 Jan;31(2):141-4.
To study the effect of ursolic acid (UA), apentacyclic triterpene acid, on MCF-7 cell apoptosis, and probable mechanism involved by detecting the expressions of caspase-3 and poly ADP-ribose polymerase(PARP) at protein level.
MCF-7 cells were cultured with different concentrations of UA. Growth inhibition of UA on MCF-7 cells was evaluated by MTT assay. Cell cycle and sub-G1 peak were performed by FCM. Morphologic changes of UA-treated cells were observed by light microscope. Apoptotic cells with condensed or fragmented nuclei were visualized by Ho 33258 staining by a fluorescence microscope (EX: U. V.). The protein expression of caspase-3 and PARP was analyzed by immunofluorescence cell staining (SABC-Cy3).
24 hours after UA treatment, inhibition of MCF-7 cell growth was concentration-dependent. The IC50 value for UA was (22.6 +/- 3.0) micromo x L(-1). Cell cycle anaysis by FCM showed that 50 micromol x L(-1) of UA arrested MCF-7 cell cycle at G0 - G1 phase. Morphological changes of MCF-7 Cells exhibited many of the hallmark features of apoptosis, including chromatin clumps and aggregation and DNA fragmentation. UA increased caspase-3 protein expression.
The results suggest that UA evokes MCF-7 cell apoptosis is correlation with the up-regulation of caspase-3. Our study indicated that UA might be a potential Chinese medical component for breast neoplasm.
研究五环三萜酸熊果酸(UA)对MCF-7细胞凋亡的影响,并通过检测半胱天冬酶-3(caspase-3)和聚ADP核糖聚合酶(PARP)在蛋白水平的表达探讨其可能机制。
用不同浓度的UA培养MCF-7细胞。采用MTT法评估UA对MCF-7细胞的生长抑制作用。通过流式细胞术(FCM)检测细胞周期及亚G1峰。用光镜观察经UA处理的细胞形态变化。用荧光显微镜(激发光:紫外光)通过Ho 33258染色观察细胞核浓缩或碎片化的凋亡细胞。采用免疫荧光细胞染色法(SABC-Cy3)分析caspase-3和PARP的蛋白表达。
UA处理24小时后,对MCF-7细胞生长的抑制呈浓度依赖性。UA的IC50值为(22.6±3.0)μmol/L。FCM细胞周期分析显示,50μmol/L的UA使MCF-7细胞周期停滞于G0 - G1期。MCF-7细胞的形态变化表现出许多凋亡的标志性特征,包括染色质团块、聚集及DNA片段化。UA增加了caspase-3蛋白表达。
结果表明,UA诱导MCF-7细胞凋亡与caspase-3的上调相关。我们的研究表明,UA可能是一种潜在的治疗乳腺肿瘤的中药成分。