大黄素抑制体内外白血病生长的实验研究

Study on emodin in growth inhibition of human promyelocytic leukemia HL-60 cell line in vitro and in vivo

目的:探讨大黄素对白血病生长的影响及其机制。方法:不同浓度大黄素(20μmol·L-1、40μmol·L-1和80μmol·L-1)作用人早幼粒白血病细胞株HL-6024h后,Cell Counting Kit-8(CCK-8)法检测细胞存活率;流式细胞术检测细胞凋亡;Western blot检测白血病细胞中Survivin的表达。建立裸鼠白血病皮下移植瘤模型,随机分为对照组、实验Ⅰ组和实验Ⅱ组3组(n=10),第3周开始分别经灌胃给予溶媒(0.1%二甲基亚砜)0.2ml/只、大黄素20mg/kg、大黄素40mg/kg,每周3次,共2周。术后第8周处死裸鼠,测量肿瘤瘤重并计算抑瘤率;免疫组织化学法检测肿瘤组织的Ki-67和Survivin的表达。结果:与对照组相比,大黄素可呈浓度依赖性抑制HL-60细胞生长,增加细胞凋亡;大黄素可明显抑制Survivin在HL-60细胞中表达,呈浓度依赖性;与对照组相比较,Ⅰ和Ⅱ组裸鼠皮下移植瘤生长均被抑制,Ki-67和Survivin表达下调,以实验Ⅱ组显著。结论:大黄素具有抑制体内外白血病生长的作用,这可能是通过抑制白血病中Survivin的表达而实现。

Objective：To investigate the effect and the mechanism of emodin in the growth inhibition of human promyelocytic leukemia HI.-60 cell line in vitro and in vivo. Method： After human promyelocytic leukemia HI. 60 cells were treated with different concentrations of emodin（20 μmol· L 1,40μmol· L 1 ,and 80 μmol · L-1 ） ,the cellular proliferation was detected by Cell Counting Kit-8（CCK-8）assay. The flow eytometry（FCM）was used to de- termine apoptosis in HL-60 cells. Western blot was used to detect the protein expression of Survivin. HL-60 cells were injected subcutaneously into nude mice to establish leukemia xenograft tumors,and the mice were randomized into 3 groups（n = 10） ： Control group, feed with 0.1 %. Dmethyl sulfoxide（DMSO） 0.2ml/D; Test group I , Emo din at dose of 20 mg/kg;Test group Ⅱ ,Emodin at dose of 20 mg/kg. All treatment lasted for two weeks,thrice per week. Eight weeks after implantation, tumor weight and inhibition rate were evaluated respectively after the mice were sacrificed. Immunohistochemistry was used to detect the positive expression of ki-67 and Survivin in the tumors. Result： Emodin induced a higher percentage of growth inhibition and apoptosis in HL-60 cell in a dose-de pendent manner when compared with the control. The expression of Survivin was down-regulated in human pro myelocytic leukemia HL-60 cells after treatment of emodin in a dose-dependent manner. In test group I and group Ⅱ , the rate of inhibition of tumor was significantly lower than that in control group,the positive expression of ki 67 and Survivin in the tumors of two test groups were significantly lower than those in control group,especially in test group Ⅱ Conclusion ： Emodin had the antitumor effect to the human promyelocytic leukemia HL-60 cells both in vitro and in vivo and the effect might be related to the down-regulation of Survivin.