摘要
目的探讨淫羊藿素致Saos2细胞凋亡及抑制其增殖的可能机制。方法采用不同质量浓度梯度淫羊藿素作用于人骨肉瘤Saos2细胞及用抗氧化剂预处理过的骨肉瘤细胞,倒置相差显微镜观察细胞形态;流式细胞仪检测不同条件下细胞活性氧的情况及凋亡情况;划痕实验检测淫羊藿素对细胞增殖迁移的影响;用终浓度为100μmol/L的氯化钴模拟骨肉瘤的缺氧环境,RT-PCR检测血管内皮生长因子(VEGF)、尿激酶型纤溶酶原激活剂受体(uPAR)、基质金属蛋白酶2(MMP2)、肾上腺髓质素(ADM)、烯醇酶-1(Enolase-1)及醛缩酶A(aldolase A)基因表达的情况。结果淫羊藿素作用后的骨肉瘤细胞活性氧水平较阴性对照组明显增高,抗氧化剂预处理组可降低细胞活性氧水平且可逆转淫羊藿素的致凋亡作用,淫羊藿素可下调VEGF、uPAR、MMP2、ADM、Enolase-1、aldolase A基因的表达。结论淫羊藿素能致骨肉瘤Saos2细胞凋亡可能跟细胞内活性氧的上调相关,增殖抑制可能与HIF1下游基因如VEGF、uPAR、MMP2、ADM、Enolase-1、aldolase A的表达下调有关。
AIM To investigate the inhibitory proliferation of icaritin and its induction of apoptosis on Saos2 cells. METHODS Saos2 cells and their pretreated cells with antioxidant were incubated with different concentra- tions of icaritin, and then the change of the cell morphology was observed by the inverted microscope. Flow cytom- etry instrument was used for the detection of reactive oxygen species （ROS） and apoptosis ratio of the cells. The scratch test for cell migration and proliferation, under the hypoxic condition simulated by cobalt dichloride （ 100 I.zmol/L） for analog of anoxic environment of osteosarcomat, and RT-PCR for measurement of VEGF, uPAR, MMP-2, ADM, Enolase-1 and aldolase A gene expression in cells were performed. RESULTS Icaritin can in- crease the ROS of Saos2 as compared with the control group, antioxidant pretreatment antagonized apoptosis ratio and relative gene expression. CONCLUSION Icaritin-inducing Saos2 apoptosis may be associated with upregula- tion of intracellular reactive oxygen, and Icaritin＇s proliferation inhibition is related to down-regulation of HIF1 downstream genes such as VEGF, uPAR, MMP2, ADM, Enolase-1 and aldolase A expression.
出处
《中成药》
CAS
CSCD
北大核心
2014年第1期4-9,共6页
Chinese Traditional Patent Medicine
作者简介
刘玉亮（1987-），男，硕士生，研究方向：中药抗骨肿瘤。Tel：18653844696，E-mail：283401832@qq．com。
通信作者：殷明（1958-），男，硕士生导师，研究方向：端粒酶基因阻抑椎间盘退变。Tel：（0791）86298917
