Guo Yong, Shan Qing-qing, Gong Yu-ping, Lin Juan, Yang Xi, Zhou Rui-qing
Department of Hematology, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, China.
Zhonghua Xue Ye Xue Za Zhi. 2012 Jun;33(6):439-43.
To investigate the anti-leukemia effect of oridonin on Ph(+) acute lymphoblastic leukemia (ALL) cell line SUP-B15.
Human Ph(+) ALL cell line was cultured in vitro. The 50% inhibition concentration (IC(50)) of oridonin against SUP-B15 cell line was examined using modified MTT 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 kinase and its downstream Akt/mTOR, Raf/MEK/ERK, STAT5 signaling pathways and the expression levels of Bcl-2 and BAX were examined by Western blot.
Oridonin inhibited the growth of SUP-B15 cell line in both time- and dose-dependent manner with the IC(50) of oridonin as (7.08 ± 1.21) µmol/L after 72 h treatment. 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 STAT5 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.
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 STAT5 signaling pathways, down-regulating the expression of Bcl-2 and up-regulating the expression of BAX.
探讨冬凌草甲素对Ph(+)急性淋巴细胞白血病（ALL）细胞系SUP-B15的抗白血病作用。
体外培养人Ph(+) ALL细胞系。采用改良MTT法检测冬凌草甲素对SUP-B15细胞系的半数抑制浓度（IC(50)）。用光学显微镜观察细胞形态学变化。通过流式细胞术分析评估药物处理后SUP-B15细胞系的凋亡率。采用蛋白质免疫印迹法检测ABL激酶及其下游Akt/mTOR、Raf/MEK/ERK、STAT5信号通路的活性水平以及Bcl-2和BAX的表达水平。
冬凌草甲素对SUP-B15细胞系的生长抑制作用呈时间和剂量依赖性，处理72 h后冬凌草甲素的IC(50)为(7.08±1.21)μmol/L。经冬凌草甲素处理的SUP-B15细胞系细胞膜变得模糊不清，部分细胞膜解体。冬凌草甲素诱导SUP-B15细胞系凋亡，0、5、10 μmol/L冬凌草甲素处理24 h后的凋亡率分别为(6.67±0.83)%、(18.30±1.79)%和(37.63±7.12)%。冬凌草甲素抑制ABL激酶及其下游Akt/mTOR、Raf/MEK/ERK和STAT5信号通路的激活，这些信号通路在SUP-B15细胞系中呈组成性激活，下调抗凋亡蛋白Bcl-2的水平，上调促凋亡蛋白Bax的表达。
冬凌草甲素通过抑制ABL激酶及其下游Akt/mTOR、Raf/MEK/ERK和STAT5信号通路的激活，下调Bcl-2的表达，上调BAX的表达，从而对Ph(+) ALL细胞系SUP-B15发挥抗白血病作用。
