芹黄素对肾癌A498细胞凋亡及HIF-1α/NF-κB信号通路的影响  

The effect of apigenin on apoptosis and HIF-1α/NF-κB signaling pathway in renal cancer A498 cells

作　　者：张莎莎[1] 黄学锋 王莉澜 章小琴 Zhang Shasha;Huang Xuefeng;Wang Lilan;Zhang Xiaoqin(Traditional Chinese Medicine Department,Huzhou First People's Hospital,Huzhou 313000,China)

机构地区：[1]湖州市第一人民医院中医科,湖州313000

出　　处：《中国医师杂志》2024年第1期70-75,共6页Journal of Chinese Physician

基　　金：浙江省教育厅一般科研项目(Y202043768)。

摘　　要：目的探讨芹黄素对肾癌A498细胞凋亡及缺氧诱导因子-1α(HIF-1α)/核因子κB(NF-κB)信号通路的影响。方法体外培养人肾癌A498细胞,分为不同浓度芹黄素(10、20、40μmol/L)组、芹黄素(40μmol/L)+HIF-1α激动剂二甲基二酰氨基乙酸(DMOG)组、HIF-1α抑制剂利非西呱(YC-1)组、对照组。采用CCK-8法和平板克隆形成实验检测细胞增殖,Hoechst 33258染色和流式细胞术检测细胞凋亡,Western blot实验检测凋亡蛋白及HIF-1α/NF-κB通路蛋白表达。结果芹黄素显著抑制A498细胞增殖,且抑制作用呈浓度依赖性(P<0.001)。与对照组比较,10、20、40μmol/L芹黄素组和YC-1组A498细胞凋亡率[(4.35±1.04)%vs(10.06±1.13)%、(18.52±2.58)%、(32.17±2.63)%、(26.94±2.41)%]、B淋巴细胞瘤-2相关蛋白(Bax)和切割型半胱氨酸天冬氨酸蛋白水解酶-3(Cleaved Caspase-3)蛋白表达量均显著升高,B淋巴细胞瘤2(Bcl-2)均显著降低(均P<0.001)。与对照组比较,10、20、40μmol/L芹黄素组A498细胞HIF-1α蛋白表达量(0.85±0.08 vs 0.63±0.06、0.31±0.03、0.16±0.02)、p-NF-κB p65/NF-κB p65比值(0.82±0.08 vs 0.51±0.05、0.30±0.03、0.13±0.01)均显著降低(均P<0.001)。与芹黄素组比较,芹黄素+DMOG组A498细胞凋亡率显著降低[(32.17±2.63)%vs(14.85±1.62)%],Bax、Cleaved Caspase-3蛋白表达量均显著降低,Bcl-2蛋白表达量均显著升高(均P<0.001)。结论芹黄素可能通过抑制HIF-1α/NF-κB信号通路活化促进肾癌A498细胞凋亡。Objective To explore the effects of apigenin on apoptosis and hypoxia inducible factor-1α(HIF-1α)/nuclear factorκB(NF-κB)signaling pathway in renal cancer A498 cells.Methods Human renal cell carcinoma A498 cells were cultured in vitro and divided into different concentrations of apigenin(10,20,40μmol/L)groups,apigenin(40μmol/L)+HIF-1αagonist dimethylenediaminoacetic acid(DMOG)group,HIF-1αinhibitor rifiximab(YC-1)group,and control group.Cell proliferation was detected using cell counting kit-8(CCK-8)assay and plate clone formation assay,apoptosis was detected using Hoechst 33258 staining and flow cytometry,and expression of apoptotic proteins and HIF-1/NF-B pathway proteins was detected using Western blot assay.Results Celery extract significantly inhibited the proliferation of A498 cells,and the inhibitory effect was concentration dependent(P<0.001).Compared with the control group,the apoptosis rates of A498 cells in the 10,20,and 40μmol/L apigenin groups and YC-1 groups were significantly increased[(4.35±1.04)%vs(10.06±1.13)%,(18.52±2.58)%,(32.17±2.63)%,(26.94±2.41)%],as well as the expression levels of B lymphocyte tumor 2 related protein(Bax)and Cleaved Caspase-3 protein,while the expression levels of B lymphocyte tumor 2(Bcl-2)were significantly reduced(all P<0.001).Compared with the control group,the HIF-1αprotein expression levels(0.85±0.08 vs 0.63±0.06,0.31±0.03,0.16±0.02)and p-NF-κB p65/NF-κB p65 ratio(0.82±0.08 vs 0.51±0.05,0.30±0.03,0.13±0.01)of A498 cells in the 10,20,and 40μmol/L apigenin groups were significantly reduced(all P<0.001).Compared with the apigenin group,the apoptosis rate of A498 cells in the apigenin+DMOG group was significantly reduced[(32.17±2.63)%vs(14.85±1.62)%],and the expression levels of Bax and Cleared Caspase-3 proteins were significantly reduced,while the expression levels of Bcl-2 proteins were significantly increased(all P<0.001).Conclusions Apigenin may promote apoptosis in renal cancer A498 cells by inhibiting the activation of the HIF-1α/NF-κ

关 键 词：肾肿瘤 细胞系 肿瘤 芹黄素 细胞凋亡 缺氧诱导因子1 Α亚基 NF-ΚB 

分 类 号：R285[医药卫生—中药学]