橄榄苦苷调节RhoA/ROCK信号通路对子宫内膜癌化疗耐药性的影响

Impact of oleuropein on chemotherapy resistance of endometrial cancer by regulating the RhoA/ROCK signaling pathway

目的观察橄榄苦苷对子宫内膜癌化疗耐药性的影响以及RhoA/ROCK信号通路发挥的作用。方法体外培养子宫内膜癌细胞Ishikawa,建立Ishikawa细胞顺铂(DDP)耐药细胞株,设对照组、DDP组、橄榄苦苷-L+DDP组、橄榄苦苷-M+DDP组、橄榄苦苷-H+DDP组;CCK8实验检测各组Ishikawa/DDP细胞增殖抑制率;流式细胞术检测各组Ishikawa/DDP细胞凋亡率;RT-PCR实验检测各组Ishikawa/DDP细胞RhoA、ROCK1 mRNA水平;Western Blot实验检测各组Ishikawa/DDP细胞RhoA、ROCK1、Cleaved-Caspase3/Caspase3、基质金属蛋白酶2(MMP2)、Ki67蛋白水平。结果与对照组比较,DDP组Ishikawa/DDP细胞各指标无显著性差异(P>0.05),橄榄苦苷-L、M、H+DDP组Ishikawa/DDP细胞RhoA、ROCK1 mRNA水平、RhoA、ROCK1、MMP2、Ki67蛋白水平均显著性降低,增殖抑制率、凋亡率、Cleaved-Caspase3/Caspase3水平显著性升高,呈剂量依赖性(P<0.05)。结论橄榄苦苷可能通过抑制RhoA/ROCK信号通路提高子宫内膜癌耐药细胞株Ishikawa/DDP对DDP的敏感性,减轻子宫内膜癌DDP化疗耐药性。

Objective To investigate the impact of oleuropein on chemotherapy resistance of endometrial cancer and the role of RhoA/ROCK signaling pathway.Methods Endometrial cancer cells Ishikawa were cultured in vitro to establish a cisplatin resistant(DDP)cell line of Ishikawa cells.They were grouped into control group,DDP group,oleuropein-L+DDP group,oleuropein-M+DDP group,and oleuropein-H+DDP group;CCK8 experiment was applied to detect the inhibition rate of Ishikawa/DDP cell proliferation in each group;flow cytometry was applied to detect the apoptosis rate of Ishikawa/DDP cells in each group;RT-PCR experiments were applied to detect the levels of RhoA and ROCK1 mRNA in Ishikawa/DDP cells in each group;Western Blot was applied to detect the levels of RhoA,ROCK1,Cleaved-Caspase3/Caspase3,MMP2,and Ki67 proteins in Ishikawa/DDP cells in each group.Results Compared with control group,there was no obvious difference in various indicators of Ishikawa/DDP cells in DDP group(P>0.05);RhoA,ROCK1 mR NA levels,RhoA,ROCK1,MMP2,and Ki67 protein levels in oleuropein-L,M,H+DDP groups were obviously reduced;the proliferation inhibition rate,apoptosis rate,and the levels of Cleaved-Caspase3/Caspase3 were obviously increased,and were dose-dependent(P<0.05).Conclusion Oleuropein may enhance the sensitivity of endometrial cancer resistant cell lines Ishikawa/DDP to DDP by inhibiting the RhoA/ROCK signaling pathway,thereby reducing DDP chemotherapy resistance in endometrial cancer.