摘要
目的:探讨黄芪甲苷对肾癌细胞增殖、凋亡的影响及其作用机制。方法:用终浓度为20μmol/L、40μmol/L、80μmol/L、160μmol/L的黄芪甲苷处理肾癌细胞A498作为不同浓度黄芪甲苷处理组,正常培养的细胞作为对照(NC)组;将anti-miR-con、anti-miR-21转染至细胞A498中,记为anti-miR-con组、anti-miR-21组;将miR-con、miR-21转染至细胞A498中再用80μmol/L黄芪甲苷处理作为HSJG+miR-con组、HSJG+miR-21组。四甲基偶氮唑盐比色法(MTT)检测细胞存活率;蛋白质印迹(Western Blot)检测裂解半胱氨酸天冬氨酸蛋白酶-3(Cleaved-caspase-3)、细胞周期蛋白D1(Cyclin D1)蛋白表达水平;流式细胞术检测细胞凋亡;实时荧光定量PCR(RT-qPCR)检测miR-21表达水平。结果:与对照组相比,不同浓度黄芪甲苷处理组肾癌细胞A498中细胞存活率显著降低,Cyclin D1表达水平显著降低,Cleaved-caspase-3表达水平显著升高,细胞凋亡率显著升高,小鼠肿块重量显著降低,miR-21表达水平显著降低(P<0.05)。miR-21低表达Cyclin D1表达水平显著降低,Cleaved-caspase-3表达水平显著升高,细胞存活率显著降低,细胞凋亡率显著升高(P<0.05)。miR-21高表达逆转了黄芪甲苷对肾癌细胞A498增殖抑制和凋亡促进的作用。结论:黄芪甲苷可抑制肾癌细胞A498增殖,促进细胞凋亡,抑制肾癌实体瘤的生长,其机制可能与miR-21表达水平有关。
Objective:To investigate the effect of astragaloside on proliferation and apoptosis of renal cell carcinoma and its mechanism.Methods:Renal cancer cell A498 was treated with astragaloside at a final concentration of 20μmol/L,40μmol/L,80μmol/L,and 160μmol/L as different concentrations of astragaloside treatment group,and normal cultured cells were used as control(NC)group.Anti-miR-con,anti-miR-21 transfected into cell A498,recorded as anti-miR-con group,anti-miR-21 group.miR-con,miR-21 were transfected into cell A498 and then used 80μmol/L astragaloside treatment as HSJG+miR-con group and HSJG+miR-21 group.MTT assay was used to detect cell viability.Western Blot assay was used to detect Cleaved-caspase-3 and Cyclin D1 protein expression.Flow cytometry was used to detect apoptosis.Real-time quantitative PCR(RT-qPCR)was used to detect miR-21 expression.Results:Compared with control group,the cell viability of the renal cell carcinoma A498 treated with different concentrations of astragaloside was significantly decreased,the expression of Cyclin D1 was significantly decreased,the expression of Cleaved-caspase-3 was significantly increased,and the apoptosis rate was significantly increased,the weight of the mouse mass was significantly reduced,miR-21 expression was significantly decreased(P<0.05).The expression of miR-21 was decreased,the expression of Cyclin D1 was significantly decreased,the expression of Cleaved-caspase-3 was significantly increased,the cell survival rate was significantly decreased,and the apoptosis rate was significantly increased(P<0.05).High expression of miR-21 reversed the effect of astragaloside on proliferation inhibition and apoptosis promotion of renal cancer cell line A498.Conclusion:Astragaloside can inhibit the proliferation of renal cell carcinoma A498 and promote cell apoptosis,inhibit the growth of solid tumors of kidney cancer.The mechanism may be related to the expression of miR-21.
作者
张悦
于学炜
于宏川
仲伟一
杨阳
Zhang Yue;Yu Xuewei;Yu Hongchuan;Zhong Weiyi;Yang Yang(Urology Department,Dalian Third People's Hospital,Liaoning Dalian 116031,China)
出处
《现代肿瘤医学》
CAS
2020年第18期3145-3150,共6页
Journal of Modern Oncology
基金
大连市卫生局科研基金项目(编号:2016-0258X)。
关键词
黄芪甲苷
MIR-21
肾癌
增殖
凋亡
astragaloside
miR-21
renal cell carcinoma
proliferation
apoptosis
