莪术醇通过调控PI3K/AKT通路促进人肝癌HepG2细胞凋亡

Curcumol promotes the apoptosis of hepatocellular carcinoma HepG2 cells by regulating PI3K/AKT pathway

目的探讨莪术醇促进人肝癌HepG2细胞凋亡的作用是否与PI3K/AKT通路有关。方法采用MTT法检测不同浓度(0、10、25、50、100μmol·L^(-1))莪术醇干预HepG2细胞24、48 h的增殖抑制率,Hoechst 33258染色法观察细胞核凋亡形态,Western blot法检测不同浓度莪术醇及PI3K/AKT通路阻断剂LY294002对HepG2细胞p-PI3K、p-AKT、Caspase-3蛋白表达的影响。结果莪术醇对HepG2细胞的增殖有明显抑制作用,呈浓度依赖性,但干预48 h对比24 h差异无统计学意义;Hoechst 33258染色发现,莪术醇干预后,同一视野下细胞数目明显减少,细胞透亮,部分细胞核碎裂,核固缩,呈现典型的凋亡形态学特征;Western blot结果显示,莪术醇能显著降低细胞p-PI3K、p-AKT蛋白的表达,上调凋亡执行蛋白Caspase-3的表达,与LY294002联用后表现出良好的协同作用,对各蛋白表达的调控较单独用药更加明显。结论莪术醇能通过抑制PI3K/AKT通路诱导人肝癌HepG2细胞凋亡。

Objective To determine whether curcumol promotes the apoptosis of human hepatoma HepG2 cells and whether it is related to PI3 K/AKT pathway. Methods MTT assay was used to detect the proliferation inhibition rate of HepG2 cells treated with curcumol at different concentrations(0, 10, 25, 50, and 100 μmol·L-1) for 24 h and 48 h. Nuclear apoptotic morphology was observed by Hoechst 33258 staining. Western blot was used to detect p-PI3 K, p-AKT, and Caspase-3 protein expression in different concentrations of curcumol and PI3 K/AKT pathway blocker LY294002. Results Curcumol significantly inhibited the proliferation of HepG2 cells in a concentration-dependent manner, but with no significant difference between the interventions at 48 h and 24 h. Hoechst 33258 staining showed that after the curcumol intervention, the number of cells in the same field was significantly reduced, the cells were translucent, part of the nuclei were fragmented, and nuclear condensation was observed, indicating typical apoptotic morphological features. Western blot showed that curcumol significantly reduced the expression of p-PI3 K and p-AKT proteins, up-regulated the expression of apoptosis executive protein Caspase-3. After combination with LY294002, curcumol showed good synergistic effect, and the regulation of each protein expression was more obvious than the use of drugs alone. Conclusion Curcumol induces the apoptosis of human hepatocellular HepG2 cells through the inhibition of PI3 K/AKT pathway.