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目的探讨PI3K/Akt通路在选择性环氧化酶2(Cox-2)抑制剂塞来昔布抗膀胱肿瘤机制中的可能作用。方法常规培养的贴壁生长的T24细胞,加入不同浓度PI3K抑制剂LY294002共培养,MTT法测定其对T24细胞增殖活性的影响;贴壁生长及悬浮培养的T24细胞,分别加入塞来昔布、PGE2、LY294002等共培养,流式细胞仪测定T24细胞凋亡率。Western blot检测塞来昔布及PGE2作用后悬浮生长的T24细胞Akt活化情况。结果塞来昔布对贴壁及悬浮培养的T24细胞均可诱导其凋亡;PGE2及LY294002均对贴壁的T24细胞增殖和凋亡无明显影响,但PGE2对悬浮生长的T24细胞可减少其凋亡,而LY294002可显著增加T24细胞的失巢凋亡率。Western blot结果提示塞来昔布显著降低而PGE2增加Akt的活化。结论塞来昔布对PI3K/Akt信号通路的主要作用是阻遏Akt的磷酸化,通过阻止Akt活化抑制T24细胞增殖并诱导其产生凋亡。
Objective To investigate the possible role of PI3K / Akt pathway in the mechanism of selective cyclooxygenase 2 (Cox-2) inhibitor celecoxib in bladder cancer. Methods The adherent T24 cells cultured in adherent culture were added with different concentrations of PI3K inhibitor LY294002 to co-culture. The effects of PI3K inhibitor LY294002 on the proliferation of T24 cells were observed by MTT assay. T24 cells adherent culture and suspension culture were added with celecoxib, PGE2, LY294002 and other co-culture, flow cytometry T24 cell apoptosis rate. Western blot was used to detect the activation of Akt in T24 cells suspended in suspension after celecoxib and PGE2 treatment. Results Celecoxib could induce apoptosis of T24 cells both in adherent and suspension culture. PGE2 and LY294002 had no significant effect on proliferation and apoptosis of adherent T24 cells, but PGE2 could reduce T24 cells in suspension Apoptosis, and LY294002 can significantly increase the ana-apoptotic rate of T24 cells. Western blot results suggest that celecoxib decreased significantly while PGE2 increased Akt activation. Conclusion The main effect of celecoxib on the PI3K / Akt signaling pathway is to suppress the phosphorylation of Akt, and to inhibit the proliferation of T24 cells and induce their apoptosis by inhibiting Akt activation.