Huang Yu-Ling, Wang Wen-Feng, Hu Jian-Da, Zheng Jun-Ting, Li Jing
Union Clinical Medical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Department of Tumor, Jinjiang Municipal Hospital, Jinjiang 362200, Fujian Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2016 Feb;24(1):8-13. doi: 10.7534/j.issn.1009-2137.2016.01.002.
To explore the effect of a new emodin derivative E11 on proliferation and apoptosis of T lymphocytic leukemia cell line Molt-4 and its possible mechanisms.
MTT method was used to plot cell growth curve. Colony culture assay was performed for studying the effect of emodin derivative E11 on colony-formation of Molt-4. The fluorescent microscopy with DAPI staining was used to examine the cell morphological changes after E11 treatment. DNA fragmentation method was used to detect the inducing effect of emodin derivative E11 on cell apoptosis. Western blot was used to determine the expressions of apoptosis-related proteins including procaspase-9, procaspase-3, PARP and PI3K/AKT, MAPK signalling pathway.
Emodin derivative E11 could strongly inhibit the growth of Molt-4 with the IC50 in 48 h at 1.381 ± 0.1552 µmol/L in dose-dependent manner. 0.1 µmol/L of E11 could inhibit cell colony formation. The typrical apopototic morphologic changes of Molt cells treated with E11 could be observed under fluorescence microscope with DAPI staining. DNA apoptotic ladder could be observed by DNA fragmentation.The expressions of procaspase -9, procaspase-3, PARP, p-MAPK, p-AKT, mTOR, p-mTOR, p-P70 and p-4BEP1 were down-regulated, while expressions of MAPK, AKT, 4EBP1 and P70 were not changed remarkably after Molt-4 were treated with E11 for 48 h.
E11 can remarkably inhibit the proliferation and induce the apoptosis of Molt-4 cells. The mechanism of apoptosis of Molt-4 cells may be related with the suppression of PI3K/AKT and MAPK signalling pathways.
探讨新型大黄素衍生物E11对T淋巴细胞白血病细胞系Molt-4增殖和凋亡的影响及其可能机制。
采用MTT法绘制细胞生长曲线。进行集落培养试验以研究大黄素衍生物E11对Molt-4集落形成的影响。采用DAPI染色荧光显微镜观察E11处理后细胞形态变化。采用DNA片段化方法检测大黄素衍生物E11对细胞凋亡的诱导作用。采用蛋白质免疫印迹法检测凋亡相关蛋白procaspase-9、procaspase-3、PARP以及PI3K/AKT、MAPK信号通路的表达。
大黄素衍生物E11能强烈抑制Molt-4细胞生长,48 h的IC50为1.381±0.1552 μmol/L,呈剂量依赖性。0.1 μmol/L的E11能抑制细胞集落形成。DAPI染色荧光显微镜下可观察到E11处理的Molt细胞典型的凋亡形态学变化。DNA片段化可观察到DNA凋亡梯带。Molt-4细胞经E11处理48 h后,procaspase -9、procaspase-3、PARP、p-MAPK、p-AKT、mTOR、p-mTOR、p-P70和p-4BEP1的表达下调,而MAPK、AKT、4EBP1和P70的表达无明显变化。
E11可显著抑制Molt-4细胞增殖并诱导其凋亡。Molt-4细胞凋亡机制可能与PI3K/AKT和MAPK信号通路的抑制有关。