Polo样激酶4抑制剂Centrinone通过调控PI3K/AKT通路影响胃癌细胞增殖和凋亡

Polo-like kinase 4 inhibitor Centrinone affects the proliferation and apoptosis of gastric cancer cells by regulating PI3K/AKT pathway

目的:探讨Polo样激酶4(PLK4)抑制剂Centrinone对胃癌细胞的增殖、迁移、凋亡的影响并进一步研究其抗肿瘤机制。方法:在TCGA数据库中分析PLK4基因在胃癌及癌旁组织的表达及对预后的影响;在人类蛋白组数据库中分析PLK4在胃癌组织中的定位;利用不同浓度(0、1、2、5μmol/L)的Centrinone处理不同胃癌细胞系,再通过CCK8法检测细胞生长情况;用transwell小室检测Centrinone对AGS细胞迁移能力的影响;用流式细胞术检测AGS细胞凋亡的情况;通过Western blot检测凋亡相关蛋白聚腺苷二磷酸-核糖聚合酶(PARP)、半胱天冬酶-3(Caspase-3)、BCL-2、BAX等的变化。结果:通过TCGA数据库分析发现PLK4在胃癌中的表达量显著高于癌旁组织(P<0.05),且与胃癌预后相关(P=0.001)。细胞生长实验显示Centrinone对AGS、BGC和SGC均有显著的生长抑制作用,其中AGS最敏感(IC50=1.386μmol/L)。细胞迁移实验提示Centrinone浓度越大,胃癌细胞迁移的数量越少(P<0.05)。流式细胞术实验结果显示Centrinone浓度越大,细胞凋亡比例越高(P<0.05)。Western blot结果显示Centrinone浓度越大,凋亡相关蛋白PARP和Caspase-3的裂解体表达量增多越明显(P<0.05),BCL-2蛋白表达量减少越明显(P<0.05),而BAX的蛋白表达量变化不明显(P>0.05),同时检测发现PI3K和AKT的磷酸化蛋白表达量显著减少(P<0.05)。结论:PLK4抑制剂Centrinone可以显著抑制胃癌细胞增殖和迁移,并诱导细胞凋亡,可能通过抑制BCL-2表达及对PI3K/AKT信号通路调控发挥作用。

Objective:To investigate the effect of Centrinone on the proliferation,migration and apoptosis of gastric cancer cells and its anti-cancer mechanism.Methods:The expression of polo-like kinase 4(PLK4)gene in gastric cancer and adjacent tissues and its effect on prognosis were analyzed in the TCGA database.The localization of PLK4 in gastric cancer tissues was analyzed in Human Proteome Atlas.Different gastric cancer cell lines were treated with different concentrations of Centrinone(0,1,2,5μmol/L),and the cell growth was detected by CCK8 method.Transwell chamber was used to detect the effect of Centrinone on the migration ability of AGS cells.The apoptosis of AGS cells was detected by flow cytometry.The apoptosis-related proteins such as poly ADP-ribose polymerase(PARP),Caspase 3,BCL-2,BAX,were detected by Western blot.Results:The expression of PLK4 in gastric cancer from TCGA database was significantly higher than that in para-cancer tissues(P<0.05),and was correlated with the prognosis of gastric cancer(P=0.001).Centrinone significantly inhibited the growth of AGS,BGC and SGC,and AGS was the most sensitive(IC50=1.386μmol/L).The migration assay showed the higher the concentration of Centrinone,the less the number of gastric cancer cells migrated(P<0.05).Flow cytometry showed that the higher the concentration of Centrinone,the higher the percentage of apoptosis(P<0.05).Western blot results showed that with the increase of Centrinone concentration,the cleavage expression of apoptosis-related proteins PARP and caspase 3 increased(P<0.05),and the expression of BCL-2 decreased(P<0.05),while the protein expression of BAX did not change significantly(P>0.05).Phosphorylated protein expression levels of PI3K and AKT were significantly decreased(P<0.05).Conclusion:PLK4 inhibitor Centrinone can significantly inhibit the proliferation,migration and induce apoptosis of gastric cancer cells,possibly by inhibiting the expression of BCL-2 and regulating the PI3K/AKT signaling pathway.