Liu Yanan, Zhu Lin, Wen Taoyu, Wan Jie, Lei Yue, Chen Hongyan
Department of Otolaryngology-Head and Neck Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Department of Otolaryngology-Head and Neck Surgery, Affiliated Hospital of South West Medical University, Luzhou 646000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 May;33(5):611-616.
Objective To study the effects of new small molecular inhibitor, forkhead domain inhibitor-6 (FDI-6), on proliferation, apoptosis, invasion and migration in human laryngeal carcinoma Hep-2 cell line and the related mechanism. Methods MTT assay was used to test the proliferation rate of Hep-2 cells before and 12, 24 hours after treated with (5, 10, 20, 40, 80) μmol/L of FDI-6. Flow cytometry (FCM) and Transwell chamber assay were respectively carried out to detect the apoptosis rate, cell invasion and migration in Hep-2 cells after treated by 10, 20 μmol/L FDI-6 for 24 hours. Real-time quantitative PCR (qRT-PCR) and Western blotting were performed to determine the mRNA and protein levels of FoxM1, Bcl-2 and BAX, respectively. Results Cell proliferation rate was inhibited by FDI-6 in a dose- and time-dependent manner. Twenty-four hours after 10, 20 μmol/L FDI-6 treatment, the apoptosis rate in Hep-2 cells was elevated and the ability of cell invasion and migration was inhibited in a dose-dependent manner. The qRT-PCR showed that there was no significant change in FoxM1 mRNA expression with or without FDI-6 treatment. Western blotting showed that the total protein level of FoxM1 was not obviously changed, but Bcl-2 was down-regulated, BAX was up-regulated. However, in the nuclear FoxM1 protein level decreased along with the ascent of FDI-6 concentration. Conclusion FDI-6 could induce cell apoptosis and inhibit cell proliferation, invasion and migration in Hep-2 cells. This may be related to the down-regulation of FoxM1 in the nucleus.
目的 研究新型小分子抑制剂叉头结构域抑制剂-6（FDI-6）对人喉癌Hep-2细胞系增殖、凋亡、侵袭和迁移的影响及其相关机制。方法 采用MTT法检测用（5、10、20、40、80）μmol/L FDI-6处理前及处理12、24小时后Hep-2细胞的增殖率。分别采用流式细胞术（FCM）和Transwell小室检测法检测10、20 μmol/L FDI-6处理24小时后Hep-2细胞的凋亡率、细胞侵袭和迁移能力。采用实时定量PCR（qRT-PCR）和蛋白质印迹法分别检测FoxM1、Bcl-2和BAX的mRNA和蛋白质水平。结果 FDI-6以剂量和时间依赖性方式抑制细胞增殖率。10、20 μmol/L FDI-6处理24小时后，Hep-2细胞凋亡率升高，细胞侵袭和迁移能力呈剂量依赖性抑制。qRT-PCR结果显示，有无FDI-6处理，FoxM1 mRNA表达均无明显变化。蛋白质印迹法显示，FoxM1总蛋白水平无明显变化，但Bcl-2下调，BAX上调。然而，核内FoxM1蛋白水平随FDI-6浓度升高而降低。结论 FDI-6可诱导Hep-2细胞凋亡，抑制其增殖、侵袭和迁移。这可能与核内FoxM1下调有关。
