TNF-α通过调控KLF4抑制口腔鳞癌细胞CAL27增殖

Effect of TNF-α on proliferation of oral squamous cell carcinoma CAL27 cells via regulating KLF4 expression

目的:探讨肿瘤坏死因子-α(TNF-α)通过上调口腔鳞癌细胞CAL27中Krüppel样因子4(KLF4)表达对细胞增殖的影响。方法:用不同浓度的TNF-α(0、2. 5、5、10、20 ng/ml)处理CAL27细胞24 h,免疫印迹试验(Western blot)检测KLF4表达变化;用20 ng/ml TNF-α处理细胞,分别于24 h、48 h、72 h用噻唑蓝(MTT)法检测细胞增殖变化;构建过表达KLF4的CAL27细胞,MTT检测其对细胞增殖的影响;MTT检测沉默KLF4(si KLF4)及si KLF4联合TNF-α处理对细胞增殖能力的影响。结果:MTT检测发现TNF-α可抑制细胞增殖;TNF-α可呈剂量依赖性地诱导CAL27细胞中KLF4表达;Western blot和qPCR检测显示成功构建过表达KLF4的CAL27细胞;过表达KLF4可抑制细胞增殖;沉默KLF4可部分恢复TNF-α对CAL27细胞增殖的抑制作用。结论:TNF-α可诱导口腔鳞癌细胞CAL27中KLF4表达增加,KLF4可能参与了TNF-α对CAL27细胞增殖的抑制作用。

Objective:To investigate the effect of TNF-αon cell proliferation by up-regulating KLF4 expression in oral squamous cell carcinoma cell line CAL27.Methods:CAL27 cells were treated with different concentrations of TNF-α(0,2.5,5,10,20 ng/ml)for 24 hours,and the expression of KLF4 protein was detected by Western blot;the cell proliferation of CAL27 treated with 20 ng/ml TNF-αwas detected by MTT assay at 24 h,48 h and 72 h,respectively;the effect of KLF4 overexpression,siKLF4 and siKLF4 combined with TNF-αon cell proliferation was detected by MTT assay.Results:The expression of KLF4 in CAL27 cells was induced by TNF-αin a dose-dependent manner;the CAL27 cells overexpressing KLF4 were constructed successfully;the proliferation of CAL27 cells was decreased by treating with 20 ng/ml TNF-αand overexpression of KLF4,respectively;silencing KLF4 could restore the inhibitory effect of TNF-αon CAL27 cells.Conclusion:TNF-αcan increase KLF4 expression in oral squamous cell carcinoma cells CAL27,and KLF4 may be involved in the inhibition of proliferation of CAL27 cells by TNF-α.