蛇床子素调节PI3K/AKT/MAPK信号通路对卵巢癌细胞增殖、凋亡、迁移和侵袭的影响

Effects of Osthole on proliferation, apoptosis, migration and invasion of ovarian cancer cells by regulating PI3K/AKT/MAPK signal pathway

目的 分析蛇床子素对卵巢癌细胞增殖、凋亡、迁移和侵袭的影响及其作用机制。方法 采用25、50、75μmol/L蛇床子素处理HO-8910细胞,以不加蛇床子素的HO-8910细胞为对照组,采用不同药物处理后分为MAPK抑制剂组(75μmol/L蛇床子素+MAPK抑制剂SB203580)及PI3K/蛋白激酶B(AKT)激活剂组(75μmol/L蛇床子素+PI3K激活剂740Y-P)。CCK-8法检测各组HO-8910细胞的增殖;流式细胞术检测各组HO-8910细胞的凋亡;Transwell实验检测各组HO-8910细胞的迁移和侵袭;Western bolt法检测各组HO-8910细胞PI3K/AKT和MAPK信号通路相关蛋白的表达水平。结果 经25、50、75μmol/L蛇床子素处理后,HO-8910细胞存活率、细胞迁移数、细胞侵袭数及p-PI3K/PI3K、p-AKT/AKT水平依次下降,细胞凋亡率、p-p38/p38水平依次升高,均呈浓度依赖性(P<0.05);MAPK抑制剂组、PI3K/AKT激活剂组HO-8910细胞存活率、细胞迁移数、细胞侵袭数均高于75μmol/L组(P<0.05),细胞凋亡率均低于75μmol/L组(P<0.05);MAPK抑制剂组HO-8910细胞p-p38/p38水平低于75μmol/L组(P<0.05),PI3K/AKT激活剂组p-PI3K/PI3K、p-AKT/AKT水平均高于75μmol/L组(P<0.05)。结论 蛇床子素可抑制HO-8910细胞增殖、迁移和侵袭,诱导其凋亡,其机制可能与抑制PI3K/AKT通路、激活p38MAPK通路有关。

Objective To analyze the effects and mechanism of Osthole on the proliferation, apoptosis, migration and invasion of ovarian cancer cells. Methods HO-8910 cells were treated with 25, 50, 75 μmol/L osthole, and other HO-8910cells without osthole treatment were used as the control group. On the basis of 75 μmol/L osthole treatment, the cells were set as MAPK inhibitor group(75 μmol/L Osthole+MAPK inhibitor SB203580) and PI3K/protein kinase B(AKT) activator group(75 μmol/L Osthole+PI3K activator 740Y-P). CCK-8 method was used to detect the proliferation of HO-8910 cells in each group. Flow cytometry was used to detect the apoptosis of HO-8910 cells in each group. Transwell experiment was used to detect the migration and invasion of HO-8910 cells in each group. Western bolt method was used to detect the expression levels of PI3K/AKT and MAPK signaling pathway related proteins in HO-8910 cells of each group. Results After treatment with 25, 50, 75 μmol/L osthole, the HO-8910 cell survival rate, cell migration number, cell invasion number, and p-PI3K/PI3K,p-AKT/AKT levels decreased sequentially, the cell apoptosis rate and p-p38/p38 level increased sequentially, all were concentration-dependent(P<0.05). The survival rate, cell migration number, and cell invasion number of HO-8910 cells in the MAPK inhibitor group and PI3K/AKT activator group were higher than those in the 75 μmol/L group(P<0.05), and the cell apoptosis rate was lower than that in the 75 μmol/L group(P<0.05). The level of p-p38/p38 of HO-8910 cells in the MAPK inhibitor group was lower than that in the 75 μmol/L group(P<0.05), and the levels of p-PI3K/PI3K and p-AKT/AKT in the PI3K/AKT activator group were higher than those in the 75 μmol/L group(P<0.05). Conclusion Osthole can inhibit the proliferation, migration and invasion of HO-8910 cells, and induce their apoptosis. The mechanism may be related to the inhibition of PI3K/AKT pathway and activation of p38MAPK pathway.