谷氨酰胺对肺腺癌A549细胞增殖、迁移及TNF-α和NF-κB表达的影响

Effects of Glutamine on Proliferation,Migration and Expression of TNF-α and NF-κB in Lung Adenocarcinoma A549 Cells

[目的]探讨谷氨酰胺（glutamine,Gln）对肺腺癌A549细胞增殖、迁移及TNF-α、NF-κB表达的影响。[方法]体外培养肺腺癌A549细胞株,CCK-8法检测不同浓度Gln（2、4、8、16、32、64mmol/L）对A549细胞的增殖影响,筛选适宜浓度。Transwell实验检测细胞迁移能力,ELISA法和Western Bolt法分别检测TNF-α、NF-κB p65蛋白表达。[结果 ]（1）Gln对A549细胞的作用与干预浓度有关,缺乏Gln时细胞生长受限;低浓度Gln（2、4、8mmol/L）对细胞增殖无明显抑制作用;高浓度Gln（16、32、64mmol/L）作用下抑制作用明显（P〈0.01）并呈时间依赖性,且32mmol/L和64mmol/L两组作用无明显差异（P〉0.05）。（2）与对照组相比,Gln组明显降低A549细胞迁移数;高浓度Gln组明显降低TNF-α、NF-κB p65蛋白的表达（P〈0.01）,但32mmol/L和64mmol/L组之间无统计学差异（TNF-α分别为14.671±0.842pg/ml、14.867±1.21pg/ml,NF-κB p65分别为0.548±0.042、0.474±0.077）（P〉0.05）。[结论 ]Gln对肺腺癌A549细胞的作用与浓度有关,32mmol/L可能为最适抑制浓度。Gln可能通过下调TNF-α和NF-κB p65蛋白表达并抑制A549细胞的迁移发挥抗肺腺癌作用。

[Objective]To investigate the effects of glutamine on proliferation,migration and expression of TNF-α and NF-κB in human lung adenocarcinoma A549 cells.[Methods]The cultured lung adenocarcinoma A549 cells were treated with glutamine（0,2,4,8,16,32,64 mmol/L）.The cell growth inhibition rate was measured with CCK-8 method;the migration ability of A549 cells was determined by Transwell assay;the expression of TNF-α was detected by ELISA method and NF-κB p65 protein was detected by Western Blot.[Results]The inhibition rates of A549 cells in glutamine 16,32 and 64 mmol/L groups were significantly higher than those in control group and glutamine 2,4,8 mmol/L groups（P0.01）,while there was no significant difference between 32 mmol/L and 64 mmol/L groups（P0.05）.Compared with the control group,the migration ability of A549 cells in glutamine 32 mmol/L group was significantly reduced;and the expressions of TNF-α and NF-κB p65 protein in in glutamine 16,32 and 64 mmol/L groups were significantly decreased（P〈0.01）,but there was no significant difference between 32 mmol/L and 64 mmol/L groups（P〉0.05）.[Conclusion]The effect of glutamine on lung adenocarcinoma A549 cells is concentration-dependent,and the optimal inhibitory concentration is 32 mmol/L.The effect of glutamine on lung adenocarcinoma cells may be related to down-regulation of TNF-α and NF-κB p65 in A549 cells.