ERK 1/2-PKM2/P53信号通路在喉癌Hep-2细胞株增殖中的作用

ERK 1/2 suppress the growth of Hep-2 cells by regulating the PKM2/P53 signaling pathway

目的:探究ERK1/2通过PKM2/P53信号通路调节喉癌Hep-2细胞株增殖的潜在机制。方法:采用细胞计数的方法统计经ERK1/2抑制剂U0126、P53抑制剂PFTα处理后0,24,48h时Hep-2细胞的细胞密度,实验随机分为空白对照组(Control)、对照组(Vehicle)、处理组(U0126或PFTα)。采用转染PKM2-siRNA或PKM2-cDNA的方法在Hep-2细胞株上抑制或过表达PKM2。采用免疫印迹(WesternBlot)的方法检测不同处理情况下ERK1/2、p-ERK1/2、P53、PKM2的蛋白水平。结果:抑制ERK1/2的磷酸化可抑制Hep-2细胞生长,抑制P53可阻断抑制ERK1/2导致的Hep-2细胞生长变缓。同时,抑制ERK1/2可下调PKM2水平,抑制或过表达PKM2可分别上调或下调P53。过表达PKM2可阻断抑制ERK1/2引起的Hep-2细胞生长变缓。结论:抑制ERK1/2的磷酸化可通过下调PKM2水平促进P53表达进而抑制Hep-2细胞生长。

O b je ctiv e: To investigate the potential mechanism of ERK 1/2 regulating Hep-2 cells prolifera-tion by modulating PKM2/P53 signaling pathway.M ethods: The cell density of Hep-2 cells was counted at 0 h, 24 h, 48 h after the cells were treated with ERK 1/2 inhibitor U0126 or P53 inhibitor PFTα by cell counting method. The cells were randomly divided into control group, vehicle group,and U0126 group or PFTα group. Inhibition or overexpression of PKM2 on Hep-2 cells was per-formed by using transfection of PKM2-siRNA or PKM2-cDNA. Western blot was used to detect the protein levels of ERK 1/2, p-ERK 1/2, P53 and PKM2 under different conditions.Results: Inhibition of ERK 1/2 inhibited the growth of Hep-2 cells, and inhibition of P53 could reverse the growth of Hep-2 cells that was inhibited by ERK 1/2. Meanwhile, inhibiting ERK 1/2 down-regulated PKM2 lev-els, and suppressing or over-expressing PKM2 could up-or down-regulate P53 expression, respec-tively. In addition, overexpression of PKM2 terminated the growth suppression of Hep-2 cells induced by ERK 1/2 inhibition.Conclusion: These results indicate that inhibition of ERK 1/2 promotes the ex-pression of P53 and subsequently inhibits the growth of Hep-2 cells by down-regulating PKM2 levels.