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目的探讨peroxynitrite(ONOO-)作用于下咽癌细胞过程中PDCD4基因表达的变化。方法以体外培养的下咽癌FaDu细胞系为实验材料,不同浓度的ONOO-作用于FaDu细胞,噻唑蓝(MTT)比色法测定细胞生存率,采用Ho.33342/PI荧光双染进行形态学观察,RT-PCR及Western-blot检测PDCD4mRNA水平及蛋白表达的变化,评价PDCD4基因在ONOO-诱导FaDu细胞凋亡过程中的作用。结果 ONOO-(50、100、200μmol/L)作用FaDu细胞24h,能够显著抑制细胞增值,并诱导细胞程序性凋亡,同时引起PDCD4mRNA水平显著上调,PDCD4蛋白表达显著增加。结论 PDCD4基因可能在ONOO-诱导下咽癌细胞生长的信号转导调节通路中起到关键作用。
Objective To investigate the changes of PDCD4 gene expression during peroxynitrite (ONOO-) treatment in hypopharyngeal carcinoma cells. Methods FaDu cell line cultured in vitro was used as experimental material. ONOO- was applied to FaDu cells in different concentrations. MTT assay was used to determine the cell viability. Ho.33342 / PI fluorescent double staining The changes of PDCD4 mRNA and protein expression were observed by RT-PCR and Western-blot, and the role of PDCD4 gene in ONOO-induced apoptosis of FaDu cells was evaluated. Results ONOO- (50, 100, 200μmol / L) treatment of FaDu cells for 24 hours significantly inhibited cell proliferation and induced programmed cell death. At the same time, PDCD4 mRNA level was significantly up-regulated and PDCD4 protein expression was significantly increased. Conclusion PDCD4 gene may play a key role in signal transduction pathway of ONOO-induced pharyngeal carcinoma cell growth.