雷公藤甲素通过miR-34b-5p/Notch1轴抑制骨肉瘤U2OS细胞增殖并诱导其铁死亡OA北大核心CSTPCD

Objective:To investigate the mechanism of triptolide(TPL)regulating Notch1 expression on ferroptosis of osteosarcoma U2OS cells via miR-34b-5p.Method:U2OS cells were routinely cultured and divided into the control group,the TPL(10 μmol/L)group,the TPL(10 μmol/L)+Fer-1(ferroptosis inhibitor,20 μmol/L)group,the miR-NC,miR-34b-5p,miR-34b-5p+Fer-1(20 μmol/L)group,the TPL(10 μmol/L)+anti-miR-34b-5p group,and the anti-miR-34b-5p+Fer-1(20 μmol/L)group.qPCR assay,CCK-8 assay,ferric ion detection reagent,DHE-fluorescent probe,and WB assay were used to detect the expression of miR-34b-5p,the proliferative ability,and the level of Fe2+,ROS levels and the expression of GPX4,SLC7A11 and Notch1 proteins,respectively.Dual luciferase reporter gene assay was used to verify the targeted binding relationship between miR-34b-5p and Notch1.Results:TPL promoted miR-34b-5p expression in U2OS cells,while Fer-1 and anti-miR-34b-5p inhibited miR-34b-5p expression(all P<0.05).TPL significantly inhibited the proliferation of U2OS cells,the expression of ferroptosis-related proteins(GPX4,SLC7A11,and Notch1 proteins),and increased the cellular Fe2+and ROS content,and Fer-1 reversed the effect of TPL on U2OS cells(all P<0.05).Overexpressing miR-34b-5p had similar effects on U2OS cells as TPL(all P<0.05).miR-34b-5p can be targeted to bind Notch1(all P<0.05).miR-34b-5p inhibitor could significantly inhibit the effect of TPL on U2OS cells(all P<0.05).Fer-1 could enhance the effect of miR-34b-5p inhibitor(all P<0.05).Conclusion:TPL inhibits the proliferative capacity and promotes ferroptosis of U2OS cells,and its mechanism may be related to the targeted regulation of Notch1 expression by miR-34a-5p.