紫草素通过PKM2/Hif-1α通路抑制视网膜母细胞瘤Y79细胞株增殖的影响及其潜在机制

Shikonin Inhibits Y79 Cell Proliferationby Inhibiting PKM2/Hif-1α Signaling

目的 探究紫草素对人视网膜母细胞瘤Y79细胞株增殖的影响及其潜在机制.方法 采用细胞计数的方法统计Y79细胞在不同处理情况下的细胞密度.采用免疫印迹方法检测Y79人视网膜母细胞瘤细胞 (Y79细胞) 在不同处理情况下的M2型丙酮酸脱氢酶 ( PKM2) 和低氧诱导因子-1α ( hypoxia inducible factor-1α, HIF-1α) 的蛋白水平.采用转染PKM2-siRNA或Hif-1α-siRNA的方法, 分别下调 Y79 细胞中的PKM2或Hif-1α水平.结果 处理紫草素可抑制Y79细胞生长, 同时, 紫草素下调Y79细胞内的PKM2和Hif-1α水平. PKM2-siRNA或Hif-1α-siRNA转染Y79 细胞下调PKM2 或Hif-1α, 可分别抑制Y79 细胞的生长.预处理抑制PKM2或Hif-1α均可阻断紫草素引起的Y79细胞生长变缓.抑制PKM2可引起Y79细胞内Hif-1α水平下调;且抑制PKM2可阻断紫草素对Y79细胞内Hif-1α水平的影响.结论 紫草素可通过抑制PKM2进而下调Hif-1α, 从而抑制Y79细胞的增殖.

Objective To investigate the effect of Shikonin on the proliferation of retinoblasto-ma Y79 cell line and its potential mechanism.Methods The cell density of Y79 cells under differ-ent treatment conditions was counted by cell counting.Western blotting was used to detect the protein levels of PKM2 and Hif-1α in Y79 cells under different treatment conditions.The level of PKM2 or Hif-1α in Y79 cells was down-regulated by transfection of PKM2-siRNA or Hif-1α-siRNA, respec-tively.Results Treatment of Shikonin inhibited the growth of Y79 cells.Meanwhile, Shikonin down-regulated the levels of PKM2 and Hif-1α in Y79 cells.PKM2-siRNA or Hif-1α-siRNA transfect-ed Y79 cells down-regulated PKM2 or Hif-1α, respectively, inhibited the growth of Y79 cells.Pretreatment inhibition of PKM2 or Hif-1α could block the slowing of Y79 cell growth induced by Shikonin.Inhibition of PKM2 caused down-regulation of Hif-1α levels in Y79 cells;and inhibi-tion of PKM2 blocked the effect of Shikonin on Hif-1α levels in Y79 cells.Conclusion Shikonin inhibits the proliferation of Y79 cells by inhibiting PKM2/Hif-1α signaling.