摘要
目的 研究雷公藤甲素对骨肉瘤细胞SaOS-2 增殖、侵袭和凋亡的影响和机制.方法 骨肉瘤细胞 SaOS-2 分成 Control 组、Triptolide 组(40、80、160 nmol·L-1 雷公藤甲素)、阳性对照组(15 mg·L-1 5-氟尿嘧啶)、Triptolide+si-NC组(转染siRNA control,160 nmol·L-1 雷公藤甲素)、Triptolide+si-BE503655 组(转染BE503655 siRNA,160 nmol·L-1 雷公藤甲素),以CCK-8 实验分析细胞变化,用流式细胞术分析细胞凋亡变化,以Transwell小室分析细胞迁移和侵袭变化,West-ern blot分析神经性钙黏附素(neural cadherin,N-cadherin)、上皮性钙黏附素(epithelical cadherin,E-cadherin)、Bcl-2 相关X蛋白(Bcl-2 associated X protein,Bax)、Bcl-2 相关X蛋白(Bcl-2 associated X protein,Bax)、基质金属蛋白酶-2(matrix metalloprotease 2,MMP-2)蛋白表达变化,qRT-PCR方法分析BE503655 表达.结果 与Control组比较,Triptolide组(40、80、160 nmol·L-1 雷公藤甲素)、阳性对照组骨肉瘤细胞存活率降低,细胞凋亡率升高,侵袭数目和迁移数目减少,细胞中Bax、E-cadherin蛋白表达水平增加,Bcl-2、MMP-2、N-cadherin蛋白表达水平降低,BE503655 表达水平升高.与Triptolide+si-NC组比较,Triptolide+si-BE503655 组骨肉瘤细胞存活率升高,细胞凋亡率降低,迁移和侵袭数目均增加,MMP-2、N-cadherin、Bcl-2 蛋白表达水平均升高,E-cadherin、Bax蛋白表达水平均降低,BE503655 水平降低.结论 雷公藤甲素通过上调BE503655 发挥抑制骨肉瘤细胞SaOS-2 增殖、迁移、侵袭、EMT和促凋亡作用.
Abstract
Objective To study the effect of triptolide on the proliferation,invasion and apoptosis of osteosarcoma cell SaOS-2.Methods SaOS-2 osteosarcoma cells were divided into Control group,Triptolide group(40,80,160 nmol·L-1 triptolide),positive control group(15 mg·L-1 5-fluorouracil),Triptolide+si-NC group(transfected siRNA control,160 nmol·L-1 triptolide),Triptolide+si-BE503655 group(transfected with BE503655 siRNA,160 nmol·L-1 triptolide),CCK-8 experiment was used to analyze cell changes,flow cytometry was used to analyze cell apoptosis,and Transwell chamber was used to analyze cell migration and invasion changes,Western blot was used to analyze N-cadherin,E-cadherin,Bax,Bcl-2,MMP-2 protein expression changes,qRT-PCR method was used to analyze the expression of BE503655.Results Compared with the control group,the survival rate of osteosarcoma cells in the Triptolide group(40,80,160 nmol·L-1 triptolide)and positive control group were decreased,the apoptosis rate was increased,and the number of invasion and migration were decreased.Bax,E-cadherin protein expression level were increased,Bcl-2,MMP-2,N-cadherin protein expression level was decreased,and BE503655 expression level was increased.Compared with the Triptolide+si-NC group,the osteosarcoma cell survival rate in the Triptolide+si-BE503655 group was increased,the apoptosis rate was decreased,the number of migration and invasion were increased,and the protein expression levels of MMP-2,N-cadherin and Bcl-2 were increased,and the expression levels of E-cadherin and Bax protein were decreased,the BE503655 level was decreased.Conclusion Triptolide exerts the effects of inhibiting proliferation,migration,invasion,EMT and promoting apoptosis of osteosarcoma cell SaOS-2 by up-regulating BE503655.
基金项目
河南省中医药科学研究专项课题(2019JDZX2080)
维普
万方数据