安罗替尼对人肾癌细胞ACHN增殖和迁移的抑制作用

Inhibitory effect of anlotinib on proliferation and migration of human renal carcinoma ACHN cells

目的观察安罗替尼(anlotinib)对人肾癌细胞增殖和迁移能力的影响,探讨其抑制肾癌细胞生长、迁移能力的机制,以及在肾癌治疗领域的临床应用前景。方法体外培养肾癌ACHN细胞,采用不同浓度(0、1.25、2.5、5和10μmol/L)的安罗替尼处理ACHN细胞24、48和72 h,MTT法检测安罗替尼对ACHN细胞增殖的影响,计算半数抑制浓度(IC50);划痕修复实验检测安罗替尼对ACHN细胞迁移能力的作用,计算迁移率;克隆形成实验检测细胞贴壁后细胞的成活能力,计算克隆数目;Western印迹法检测安罗替尼对PI3K/AKT信号通路相关蛋白表达和活性的影响,用灰度分析方法计算其灰度值。结果安罗替尼能显著抑制ACHN细胞增殖,具有明显的浓度依赖性;与对照组相比,安罗替尼处理后,能明显抑制ACHN细胞的克隆形成、迁移和信号通路相关蛋白的活化(P<0.05)。结论安罗替尼能显著抑制人肾癌ACHN细胞的增殖、克隆形成和迁移能力,其机制与PI3K信号通路的抑制相关。

Objective To evaluate the effect of anlotinib on the proliferation and migration of human renal carcinoma ACHN cells,and investigate the possible mechanism of anlotinib and potential clinical applications of anlotinib in the treatment of renal cancer. Methods ACHN cells were cultured in vitro and treated with anlotinib at different concentrations(0,1.25,2.5,5 and 10 μmol/L)for 24,48 and 72 h respectively. The effect of anlotinib on ACHN cell proliferation was assayed by the MTT method to determine the IC50 of anlotinib. The effect of anlotinib on ACHN cell migration was tested by the wound healing assay to obtain related migration rates. The colony formation assay was used to detect the viability of ACHN cells after cell attachment, and then the number of formed ACHN cell colonies were counted. Western blotting was used to investigate the expressions of related proteins of the PI3K/AKT signaling pathways,and gray degree analysis was performed to obtain related gray degree values. Results Anlotinib significantly inhibited the proliferation of ACHN cells in both time-and dose-dependent manners. Compared with the control groups,anlotinib significantly inhibited ACHN cell colony formation,cell migration and activation of the PI3K signaling pathway related proteins(P<0.05).Conclusion Anlotinib can significantly inhibit the proliferation,colony formation and migration of human renal carcinoma ACHN cells,and the possible mechanism is related to the inhibition of the PI3K signaling pathway by anlotinib.