Role of mTORC1 in the process of promoting breast cancer induced by arachidonic acid metabolites

LIU Miao; ZHANG Yue; HE Min-hong; ZHOU Ling; ZHENG Hang

doi:10.3969/j.issn.1002-266X.2014.14.009

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 mTORC 1 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 prolifera-tion 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), re-spectively .The grey value of P-S6 of HETE group was higher than that of the control group , whereas the grey value of P-S6 of 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.