Zhou Honghui, Zhao Haibo, Liu Hui, Xu Xiang, Dong Xiaoli, Zhao Enfeng
Department of Obstetrics and Gynecology, Chinese PLA General Hospital, No.28, Fuxing Rd, Beijing 100853,China.
J BUON. 2018 Nov-Dec;23(6):1732-1738.
To investigate the influence of carboplatin on the proliferation and apoptosis of ovarian cancer cells through mTOR/P70S6K signaling pathway.
The mRNA and protein expressions were detected via Western blotting and RT-PCR to study whether the mTOR/p70S6K signaling pathway was activated in OVCAR-3 and Caov-3 ovarian cancer cell lines. After cells were treated with different concentrations of carboplatin, the mRNA and protein expressions of mTOR, p70S6K and 4E-BP1 were detected via RT-PCR and Western blotting. OVCAR-3 cells were treated with 20 and 50 μM carboplatin for 4 hrs, and then apoptosis was analyzed and assessed. OVCAR-3 cells were treated with different concentrations of carboplatin (20, 50, 100, 150 and 200 μM) for 24 and 48 hrs, respectively.
The mTOR signaling pathway was activated in OVCAR-3 and Caov-3 ovarian cancer cell lines. The mRNA level of mTOR in Caov-3 cells was higher, but that of p70S6K was lower. Carboplatin significantly reduced the mRNA expression of mTOR (p<0.01), whereas the mRNA expressions of p70S6K and 4E-BP1 in carboplatin-treated cells were increased in a dose-dependent manner (p<0.01). Carboplatin inhibited the mTOR protein expression in a dose-dependent manner (p<0.01). The proliferation of OVCAR-3 cells exposed to carboplatin was reduced compared with that of untreated cells (p<0.01), and the inhibitory effect of carboplatin on the proliferation of OVCAR-3 cells was time- and dose-dependent.
The mTOR/p70S6K pathway was activated in ovarian cancer. Carboplatin could rapidly inhibit the expression of mTOR, and the phosphorylation of its major downstream effectors p70S6K and 4E-binding protein 1 (4E-BP1) arrested cells in G0/G1 phase and induced ovarian cancer cell apoptosis.
通过mTOR/P70S6K信号通路研究卡铂对卵巢癌细胞增殖和凋亡的影响。
通过蛋白质免疫印迹法（Western blotting）和逆转录聚合酶链反应（RT-PCR）检测mRNA和蛋白质表达，以研究mTOR/p70S6K信号通路在OVCAR-3和Caov-3卵巢癌细胞系中是否被激活。用不同浓度的卡铂处理细胞后，通过RT-PCR和蛋白质免疫印迹法检测mTOR、p70S6K和4E-BP1的mRNA和蛋白质表达。用20和50μM卡铂处理OVCAR-3细胞4小时，然后分析和评估细胞凋亡情况。分别用不同浓度（20、50、100、150和200μM）的卡铂处理OVCAR-3细胞24小时和48小时。
mTOR信号通路在OVCAR-3和Caov-3卵巢癌细胞系中被激活。Caov-3细胞中mTOR的mRNA水平较高，但p70S6K的mRNA水平较低。卡铂显著降低了mTOR的mRNA表达（p<0.01），而在卡铂处理的细胞中，p70S6K和4E-BP1的mRNA表达呈剂量依赖性增加（p<0.01）。卡铂以剂量依赖性方式抑制mTOR蛋白表达（p<0.01）。与未处理的细胞相比，暴露于卡铂的OVCAR-3细胞的增殖减少（p<0.01），卡铂对OVCAR-3细胞增殖的抑制作用具有时间和剂量依赖性。
mTOR/p70S6K通路在卵巢癌中被激活。卡铂可迅速抑制mTOR的表达，其主要下游效应分子p70S6K和4E结合蛋白1（4E-BP1）的磷酸化使细胞停滞在G0/G1期并诱导卵巢癌细胞凋亡。
