CPI203抑制肾癌ACHN细胞增殖及其机制

Antitumor activity of CPI203 in renal cell carcinoma ACHN cells and its mechanism

目的研究CPI203杀伤人肾癌ACHN细胞的能力及潜在机制。方法不同浓度(0、0.1、0.5、1、5μmol/L)CPI203药物作用于ACHN细胞24、48 h后,采用CCK8试剂盒检测CPI203抑制ACHN细胞增殖效果;采用流式细胞术分析CPI203对ACHN细胞凋亡及细胞周期的影响;划痕实验观察CPI203影响ACHN细胞迁移及侵袭能力;克隆形成实验检测CPI203抑制ACHN细胞克隆集落形成能力;荧光定量PCR法及免疫印迹法分别分析CPI203作用于ACHN细胞后, MYC、 NOXA、 AKT、 ERK、CyclinD1和GSK3β的表达量变化情况。结果 CPI203明显抑制肾癌细胞株ACHN细胞增殖,并呈时间及浓度依赖性(P<0.05);CPI203药物可促进ACHN细胞凋亡并抑制细胞生长周期(P<0.05);CPI203呈浓度依赖性地降低ACHN细胞的克隆形成能力及迁移能力(P<0.05);CPI203降低肾癌ACHN细胞中MYC、NOXA、AKT、ERK、CyclinD1和GSK3β表达量减少(P<0.05)。结论CPI203可抑制ACHN细胞增殖,促进肾癌细胞凋亡并抑制其生长周期进展,明显抑制癌细胞迁移及克隆形成能力,其作用机制与CPI203影响肾癌ACHN细胞中MYC、NOXA、AKT、ERK、CyclinD1和GSK3β表达量有关。

Objective To explore the ability of CPI203 killing renal cell carcinoma ACHN cells and its potential mechanism.Methods ACHN was treated with various concentrations (0,0.1,0.5,1 and 5 μmol/L) of CPI203 for 24 and 48 h.The effect of CPI203 on ACHN cell proliferation was assessed by CCK8.Flow cytometry was applied to examine the effect of CPI203 on ACHN cell cycle and apoptosis.Wound healing assay and colony forming assay were applied to evaluate the capacity of cell migration and colony formation,respectively.Quantitative Real-time PCR and Western blotting were applied to evaluate the mRNA and protein expression of MYC,NOXA,AKT,ERK,CyclinD1 and GSK3β.Results CPI203 inhibited the growth of ACHN cells.CPI203 induced both apoptosis and cell cycle arrest of ACHN cells in a dose-dependent manner.The migration and colony forming ability were also inhibited by CPI203 in a dose-dependent manner.CPI203 decreased relating genes expression,such as MYC,NOXA,AKT,ERK,CyclinD1 and GSK3β.Conclusion CPI203 has significant antitumor effect against renal tumor cells via inhibition of MYC,NOXA,AKT,ERK,CyclinD1 and GSK3β.