摘要
目的研究Stat1蛋白对宫颈癌Hela细胞生长、增殖及顺铂敏感性的影响及其作用机制。方法采用蛋白印迹法(Western blot)检测不同浓度DDP处理Hela细胞后Stat1蛋白表达差异;转染Stat1-si RNA后,通过Western blot验证干扰效果;应用四甲基偶氮唑蓝(MTT)法和5-溴脱氧尿嘧啶核苷(Brd U)观察转染Stat1-si RNA后Hela细胞对DDP敏感性变化,并检测Stat1相关细胞因子c-Myc的表达变化。结果随着DDP浓度的增加,Hela细胞中Stat1蛋白的表达量逐渐增加,DDP浓度为5 mg/L时,Stat1蛋白表达上调1.5倍,DDP浓度为10 mg/L时,Stat1蛋白表达上调近2倍;特异性Stat1-si RNA使Hela细胞中Stat1的表达下调70%,促进细胞生长和增殖;Stat1-si RNA可以降低DDP对Hela细胞生长和增殖的抑制效应,削弱DDP对c-Myc的抑制作用。结论 Hela细胞中c-Myc是STAT1发挥抑癌作用的靶点基因之一;STAT1/c-Myc通路介导了DDP抑制Hela细胞生长、增殖的作用;STAT1可增强Hela细胞对DDP的敏感性。
Objective To investigate the role of Stat1 gene in the proliferation and chemotherapeutic sensitivity of cervical cancer He La cells. Methods The protein expression of Stat1 in the Hela cells exposed to gradient concentrations of cisplatin（DDP） was detected by Western blotting with or without small interfering RNA（si RNA）- mediated Stat1 gene silencing. The effect of Sata1 silencing on the sensitivity to DDP and cell proliferation of the cells was tested using MTT assay and Brd U assay,and the expression of c-Myc was detected by Western blotting in the cells treated with si RNA and DDP. Results The expression of Stat1 in Hela cells exposed to DDP increased with the DDP concentrations, reaching 1.5 folds of the baseline at a DDP concentration of 5 mg/L and 2 folds at 10 mg/L. Stat1-si RNA effectively reduced Stat1 expression in Hela cells, promoted the cell proliferation, and enhanced the expression of c-Myc; DDP inhibited the cell growth and down-regulated c-Myc expression.Stat1-si RNA rescued DDP-induced inhibition of cell growth and c-Myc down-regulation. Conclusion The expression of Stat1 is associated with DDP sensitivity in cervical cancer cells, and Stat1 silencing can increase the sensitivity to DDP and c- Myc expression of the cells.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2015年第1期88-92,共5页
Journal of Southern Medical University
基金
国家自然科学青年科学基金(81402270)
湖南省科技厅重点项目(2012FJ2014)~~
