醌茜素对人肝癌Huh7细胞的凋亡诱导作用及其抗肿瘤机制

发布时间：2018-02-03 13:29

本文关键词： 醌茜素肝癌细胞凋亡 Akt信号通路活性氧　出处：《肿瘤防治研究》2017年07期 　论文类型：期刊论文

【摘要】：目的探讨醌茜素诱导人肝癌Huh7细胞凋亡的机制。方法 MTT法测定醌茜素对人肝癌Hep G2、Hep3B和Huh7细胞的杀伤作用;倒置显微镜下观察醌茜素处理后Huh7细胞形态学变化;Annexin V-FITC/PI双染法及流式细胞术检测其对肝癌Huh7细胞的诱导凋亡作用;蛋白质免疫印迹法检测细胞凋亡相关蛋白的表达量变化;流式细胞术检测醌茜素处理后细胞内ROS的表达水平。结果与对照组(0 h)相比醌茜素能够明显抑制人肝癌细胞的存活率(P=0.0204)。经醌茜素处理后的实验组Huh7细胞呈现经典的细胞皱缩、核固缩等现象。实验组(24 h)Huh7细胞凋亡率为50.63%,明显高于对照组(0 h)(15.58%)(P=0.0001)。共同处理Akt抑制剂和醌茜素后,促凋亡蛋白Bad和cleaved-caspase-3表达量均逐渐增加,抗凋亡蛋白磷酸化的Akt和Bcl-2均逐渐减少(P0.05)。醌茜素处理后,可增加Huh7细胞内活性氧的水平。结论醌茜素对人肝癌Huh7细胞具有杀伤作用,且呈浓度依赖性。其机制可能是醌茜素抑制细胞内Akt的表达及磷酸化水平或诱导活性氧的产生,进而促进Huh7细胞的凋亡。
[Abstract]:Objective to investigate the mechanism of apoptosis induced by alizarin in human hepatocellular carcinoma (Huh7) cells. Methods MTT assay was used to determine the killing effect of alizarin on Hep G _ 2 Hep3B and Huh7 cells. The morphologic changes of Huh7 cells treated with alizarin quinone were observed under inverted microscope. Annexin V-FITC / Pi double staining and flow cytometry were used to detect the apoptosis of Huh7 cells. The expression of apoptosis-related protein was detected by Western blot. Flow cytometry was used to detect the expression of ROS in the cells treated with alizarin. Results compared with the control group for 0 h, alizarin could significantly inhibit the survival rate of human hepatoma cells. The Huh7 cells of the experimental group treated with alizarin showed classic cell shrinkage. The apoptosis rate of Huh7 cells in the experimental group was 50.63% at 24 h. It was significantly higher than that of the control group at 0 h after treatment with Akt inhibitor and quinoid alizarin. The expression of apoptotic protein Bad and cleaved-caspase-3 increased gradually. The phosphorylated Akt and Bcl-2 of anti-apoptotic protein decreased gradually (P 0.05). Conclusion alizarin can kill human hepatoma Huh7 cells. The mechanism may be that alizarin inhibits the expression of Akt and phosphorylation or induces the production of reactive oxygen species (Ros), which may promote the apoptosis of Huh7 cells.
【作者单位】：黑龙江八一农垦大学生命科学技术学院;哈尔滨医科大学第五医院妇产科;
【基金】：黑龙江省自然科学基金(LC2015036) 黑龙江省大学生创新创业训练计划项目(201710223001) 黑龙江八一农垦大学校内培育课题(XA2015-04);黑龙江八一农垦大学研究生创新科研项目(YJSCX2017-Y72)
【分类号】：R735.7
【正文快照】： 0引言肝癌是最常见的恶性肿瘤之一,其致死率仅次于胃癌、食管癌,在世界范围内排第三位[1-2]。现今有效治疗肝癌的方法是进行手术切除或肝移植,但临床上能得到有效治疗的患者仅占30%~40%,因此,寻找有效治疗肝癌的药物极为重要[2-4]。醌茜素(quinalizarin,化学名1,2,5,8-四羟基

【参考文献】