摘要
目的:探讨芹菜素对人肺癌H1975细胞增殖、凋亡和转移及胰岛素样生长因子1受体(IGF-1R)/蛋白激酶B(Akt)信号通路的影响。方法:设H1975细胞组、氟尿嘧啶组(100μg·ml-1)、芹菜素低(100μg·ml-1)、高(200μg·ml-1)剂量组,以上各组每孔设6个平行样,培养72h。MTT法、结晶紫染色测定细胞增殖、克隆形成数目,Transwell细胞培养室评估细胞迁移,流式细胞仪测定凋亡水平,实时荧光定量逆转录-聚合酶链反应(RT-PCR)法及蛋白免疫印迹(WEST-BLOT)法检测IGF-1R、Akt基因和蛋白表达水平。结果:与H1975细胞组比较,氟尿嘧啶组、芹菜素低、高剂量组的光密度(OD)值、存活率、细胞克隆形成数目、穿膜数、IGF-1R、Akt mRNA和蛋白降低,凋亡率升高(P<0.05)。与氟尿嘧啶组比较,芹菜素低剂量组的光密度(OD)值、存活率、细胞克隆形成数目、穿膜数、IGF-1R、Akt mRNA和蛋白升高,凋亡率降低(P<0.05);芹菜素高剂量组OD值、存活率、细胞克隆形成数目、穿膜数、IGF-1R、Akt mRNA和蛋白降低,凋亡率升高(P<0.05)。与芹菜素低剂量组比较,芹菜素高剂量组的OD值、存活率、细胞克隆形成数目、穿膜数、IGF-1R、Akt mRNA和蛋白降低,凋亡率升高(P<0.05)。结论:芹菜素能明显抑制人肺癌H1975细胞增殖、凋亡和转移;其机制与芹菜素抑制IGF-1R、Akt mRNA和蛋白的表达,进而抑制了IGF-1R/Akt信号通路的激活有关。
Objective: To investigate the effects of apigenin on the proliferation,apoptosis and metastasis of human lung cancer H1975 cells and insulin-like growth factor 1 receptor( IGF-1R)/protein kinase B( Akt) signaling pathway. Methods: H1975 cell group,5-fluorouracil group( 100 μg·ml-1),low and high dose apigenin groups( 100 μg·ml-1,200 μg·ml-1) were set up. Six parallel samples were established in each hole for 72 hours. After culture,MTT and crystal violet staining were used to determine the cell proliferation and clone formation. Transwell cell culture chamber was used to evaluate the cell migration. Flow cytometry was used to detect the apoptotic level. RT-PCR and Western blot were used to detect the expressions of IGF-1R and Akt mRNA and protein. Results: Compared with those in H1975 cell group,the OD value,survival rate,number of cell clones,number of membrane penetration,IGF-1R,Akt mRNA and protein in 5-fluorouracil group,low dose apigenin group and high dose apigenin group were decreased,and the apoptosis rate was increased( P<0.05). Compared with those in 5-fluorouracil group,the OD value,survival rate,number of cell clones,number of membrane penetration,IGF-1 R,Akt mRNA and protein in low dose apigenin group were increased,and the apoptosis rate was decreased( P<0.05);and the OD value,survival rate,number of cell clone formation,number of membrane penetration,IGF-1 R,Akt mRNA and protein of high dose apigenin group were decreased,and the apoptosis rate was increased( P< 0.05).Compared with those in low dose apigenin group,the OD value,survival rate,number of cell clone formation,number of membrane penetration,number of IGF-1R,Akt mRNA and protein of high dose apigenin group were decreased,and the apoptosis rate was increased( P<0.05).Conclusion: Apigenin can significantly inhibit the proliferation and metastasis of human lung cancer H1975 cells,and its mechanism is related to the inhibition of the expressions of IGF-1R,Akt mRNA and protein by apigenin,which further inhibits the activation of IGF-1R/Akt signaling pathway.
作者
黄茂
王星钧
孔一名
Huang Mao;Wang Xingjun;Kong Yiming(Department of Respiratory and Critical Care Medicine,Taizhou First People's Hospital,Zhejiang Taizhou 318020,China)
出处
《中国药师》
CAS
2020年第12期2384-2389,共6页
China Pharmacist
