【摘要】 目的探讨花生四烯酸(AA)代谢产物5-羟二十烷四烯酸(HETE)、12-HETE促进乳腺癌发生过程中雷帕霉素靶蛋白1(mTOR)信号通路的作用。方法将体外培养乳腺癌MCF-7细胞随机分为4组,对照组加入0.1%DMSO,HETE组加入5-HETE及12-HETE 15μmol/L,Rap组加入mTORC1抑制剂雷帕霉素(Rap)100μmol/L,HETE+Rap组加入Rap 100μmol/L及5-HETE+12-HETE 15μmol/L。CCK-8法检测各组细胞增殖率,免疫印迹法检测各组细胞mTORC1信号通路下游蛋白P-S6活性,细胞划痕实验检测各组细胞迁移能力。结果细胞增殖率HETE组高于对照组,HETE+Rap组低于HETE组(P均<0.05)。P-S6蛋白灰度值HETE组高于对照组,HETE+Rap组低于HETE组(P均<0.01)。MCF-7细胞迁移距离HETE组大于对照组(P<0.01)。HETE+Rap组小于HETE组(P<0.05)。结论 mTORC1信号通路参与了AA代谢产物乳腺癌发生的过程。更多还原

【Abstract】 Objective To investigate the role of mammalian target of rapamycin( mTOR) complex 1 in the process of promoting breast cancer induced by arachidonic acid( AA) metabolites 5,12- hydroxy-eicosatetraenoic acid( HETE).Methods Breast cancer MCF-7 cells were cultured and then were divided into four groups. The control group was cultured with 0. 1% DMSO. HETE group was cultured with 15 μmol /L 5-HETE and 12-HETE. Rap group was cultured with 100μmol/L mTORC1 inhibitor rapamycin( Rap). HETE + Rap group was cultured with Rap 100 μmol/L and 15 μmol/L 5-HETE + 12-HETE. CCK-8 assay was used to detect the rate of cell proliferation. Western blotting was used to detect the activity change of the downstream protein mTORC1 signaling pathway P-S6 protein. Wound healing was used to detect the migration activity. Results The cell proliferation rates of the control group,HETE group,Rap group,and HETE + Rap group was( 0. 85 ± 0. 01),( 1. 20 ± 0. 13),( 0. 53 ± 0. 08),and( 0. 54 ± 0. 01),respectively. The cell proliferation rate of the HETE group was higher than that of the control group,whereas the cell proliferation rate of HETE + Rap group was lower than that of HETE group( all P < 0. 05). The grey value of P-S6 of the control group,HETE group,Rap group,and HETE + Rap group was( 0. 85 ± 0. 01),( 1. 20 ± 0. 13),( 0. 53 ± 0. 08),and( 0. 54 ± 0. 01),respectively. The grey value of P-S6 of HETE group was higher than that of the control group,whereas the grey value of P-S6of HETE + Rap group lower than that of the HETE group( all P < 0. 01). The MCF-7 cell migration distance in the HETE group was longer than that of the control group( P < 0. 01),whereas MCF-7 cell migration distance in the HETE + Rap group was shorter than that of the HETE group( P < 0. 05). Conclusion mTORC1 is involved in the process of promoting breast cancer induced by 5,12-HETE.更多还原