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目的研究硫链丝菌肽(thiostrepton,TST)对人喉癌Hep-2细胞株增殖能力及FoxM1表达的影响。方法用MTT法测定TST作用于人喉癌Hep-2细胞48 h的增殖抑制率;通过流式细胞仪(FCM)检测不同浓度TST分别作用于Hep-2细胞48 h后对其细胞周期动力学的影响;采用免疫细胞化学法、RT-PCR和Western blot检测Hep-2细胞中FoxM1mRNA及蛋白表达,以及不同浓度TST作用于Hep-2细胞48 h FoxM1蛋白表达的变化。结果不同浓度(1、2、4、8、12、16μmol/L)TST作用于Hep-2细胞48 h后的增殖抑制率分别为5.27%、9.43%、20.25%、41.61%、67.96%、96.95%;浓度分别为2、3μmol/L的TST作用于Hep-2细胞48 h后,均使Hep-2细胞停滞于G1/G0期(P<0.05),且呈剂量依赖性(P<0.05);同时将TST作用于Hep-2细胞48 h后,经免疫细胞化学法、RT-PCR和Western blot均检测到FoxM1 mRNA及蛋白在Hep-2细胞中表达显著降低(P<0.05)。结论 TST对人喉癌Hep-2细胞株具有明显的增殖抑制作用,其可能机制是与降低肿瘤细胞中FoxM1蛋白表达有关。
Objective To investigate the effect of thiostrepton (TST) on proliferation and FoxM1 expression in human laryngeal carcinoma Hep-2 cell line. Methods The inhibitory rate of TST on the proliferation of human laryngeal carcinoma Hep-2 cells was determined by MTT assay. The cell cycle kinetics of Hep-2 cells treated with different concentrations of TST were detected by flow cytometry (FCM) The expressions of FoxM1 mRNA and protein in Hep-2 cells were detected by immunocytochemistry, RT-PCR and Western blot, and the changes of FoxM1 protein expression in Hep-2 cells treated with different concentrations of TST for 48 h. Results The proliferation inhibitory rates of TST treated with different concentrations of TST for 48 h were 5.27%, 9.43%, 20.25%, 41.61%, 67.96% and 96.95%, respectively %, Respectively. Both Hep-2 cells and Hep-2 cells were arrested in G1 / G0 phase (P <0.05) at a dose of 2,3μmol / L for 48 hours, At the same time, the expression of FoxM1 mRNA and protein in Hep-2 cells was significantly decreased by immunocytochemistry, RT-PCR and Western blot 48 h after TST treatment (P <0.05). Conclusion TST has a significant inhibitory effect on human laryngeal carcinoma Hep-2 cell line, which may be related to the decrease of FoxM1 protein expression in tumor cells.