摘要
目的探讨冬凌草甲素对Ph染色体阳性（Pb＋）急性淋巴细胞白血病（ALL）的抗白血病效应及其机制。方法以Ph＋ALL细胞株SUP—B15为研究对象，应用改良MTT法测定冬凌草甲素对SUP—B15细胞增殖的影响，计算72h的半数抑制浓度（IC50）；光学显微镜下观察冬凌草甲素处理后SUP—B15细胞的形态学变化；用流式细胞术检测药物作用前后SUP—B15细胞的凋亡率；以K562细胞为对照，应用Westernblot法比较药物作用前后SUP—B15细胞Akt／mTOR、Raf／MEK／ERK、STAT5信号转导通路活化水平，以及凋亡调节蛋白Bcl-2和Bax的变化。结果①冬凌草甲素呈浓度及时间依赖地抑制SUP—B15细胞增殖，作用72h的IC50值为（7．08±1．21）μmol／L；②冬凌草甲素处理24h后光学显微镜下SUP—B15细胞边界不清晰，部分细胞崩解；③0、5、10μmol／L冬凌草甲素作用24h后，SUP—B15细胞凋亡百分率分别为（6．67±0．83）％、（18．30±1．79）％、（37．63±7．12）％；④冬凌草甲素明显抑制SUP-B15细胞ABL酪氨酸激酶及其下游Akt／mTOR、Raf／MEK／ERK、STAT5多条信号转导通路的活化；下调SUP．B15细胞抗凋亡蛋白Bcl-2的表达，上调促凋亡蛋白Bax的表达。结论冬凌草甲素通过抑制ABL酪氨酸激酶及其下游Akt／mTOR、Raf／MEK／ERK、STAT5信号转导，以及上调促凋亡蛋白Bax、下调抗凋亡蛋白Bcl-2而发挥抗Ph＋ALL作用。
Objective To investigate the anti-leukemia effect of oridonin on Ph＋ acute lymphoblastic leukemia（ALL） cell line SUP-B15. Methods Human Ph＋ ALL cell line was cultured in vitro. The 50% inhibition concentration （1C50） of oridonin against SUP-B15 cell line was examined using modified MTr assay. The cellular morphologic changes were observed using a light microscope. The percent of apoptosis of SUP-B15 cell line after drug treatment was evaluated by flow cytometric analysis. The active levels of ABL ki- nase and its downstream Akt/mTOR, Raf/MEK/ERK, STATS signaling pathways and the expression levels of Bcl-2 and BAX were examined by Western blot. Results Oridonin inhibited the growth of SUP-B15 cell line in both time- and dose-dependent manner with the ICs0 of oridonin as （7.08 ±1.21 ）μmol/L after 72 h treat- ment. The cellular membrane of SUP-B15 cell line treated with oridonin became unsharp, some of them disintegrated. Oridonin induced apoptosis in SUP-B15 cell line with the apoptosis rates following 0, 5, 10 μmol/L oridonin treatment for 24 h were （6.67 ± 0.83 ） %, （ 18.30 ± 1.79 ） % and （37.63± 7.12 ） %, respectively. Oridonin inhibited activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STATS signaling pathways,which were constitutively activated in SUP-B15 cell line, down-regulated the level of anti- apoptotic protein Bcl-2 and up-regulated the expression of pro-apoptotic protein Bax. Conclusion Oridonin exerted anti-leukemia effect in Ph＋ ALL cell line SUP-B15 by inhibiting the activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STATS signaling pathways, down-regulating the expression of Bcl-2 and up-regulating the expression of BAX.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2012年第6期439-443,共5页
Chinese Journal of Hematology
基金
国家自然科学基金（30770912）
四川省科技厅社会公益项目（2008SZ0017）
教育部回国留学人员启动基金（20071108-18-4）
关键词
白血病
淋巴细胞
急性
费城染色体
细胞增殖
细胞凋亡
冬凌草素
细胞内信号肽和蛋白质类
Leukemia,lymphocytic, acute
Philadelphia chromosome
Cell proliferation
Apoptosis
Rubescensine
Intracellular signaling peptides and proteins
