二氧化硒对肺癌细胞株生长及c-fos蛋白表达的影响

Influence of the growth and c-fos expression in the GLC-82 cell line induced by selenium dioxide

目的通过研究不同浓度的二氧化硒(SeO_2)对GLC-82肺癌细胞株的生长及c-fos蛋白表达的影响,初步探讨SeO_2诱导肺癌细胞凋亡的作用机制。方法实验采用MTT比色的方法来观察不同浓度的SeO_2作用于GLC-82细胞24～96 h后,对细胞增殖和存活的影响;c-fos蛋白的检测方法为:把GLC-82细胞分别接种于6孔板中,培养爬片24h后加入SeO_2,使SeO_2的浓度分别为3、10、30μmol/L,保留1个孔作为空白对照。分别在加入SeO_2后1、6、12和24 h捞片,固定后进行SP法的c-fos蛋白免疫组化染色。在光镜10×40倍视野下,应用CMIAS-Ⅱ真彩色图像分析仪对阳性细胞的图像学参数进行分析,结果经Spss 8.0统计软件统计分析。结果GLC-82细胞在3、10、30μmol/L的SeO_2作用下,24～96 h内生长受到明显的抑制;SeO_2在早期1 h即可引起c-fos蛋白表达增强(P>0.05),随时间的延长,c-fos蛋白表达增强逐渐加强;在6～12 h,GLC-82细胞c-fos均呈明显的高表达(P<0.05和P<021),24 h仍维持较高表达水平,并且高剂量组(30μmol/L)c-fos蛋白的表达强度显著高于低剂量组(3μmol/L,P<0.05)。结论SeO_2对肺癌细胞GLC-82有生长抑制作用,并可引起GLC-82细胞c-fos蛋白的高表达,提示c-fos基因参与SeO_2对肺癌细胞凋亡的调节过程,且具有一定的剂量效应关系。

Objective This study was designed to observe the effects of selenium dioxide （SeO2） on the growth and c-fos expression in lung cancer GLC-82 cell line. Methods MTT assay was used to observe the inhibition of growth of GLC-82 cells treated with SeO2. After exposed to SeO2 for 1, 6, 12 and 24 h at different concentrations （3, 10 and 30μmol/L）, immunohistochemical method was used to investigate the expression of c-fos, and stereological analysis for c-fos positive cells was performed by CMIAS-II image analysis system at a magnification of 400. Results After treated with SeO2 for 24-96 h, cell proliferation and viability were markedly inhibited. The expression of c-los was up-regulated at 1 h （P 〉0.05）, and there were significant differences at 6-12 h （P 〈0.05 and P 〈0.01）. The level of c-los expression in group of 30 μmol/L was higher than that in group of 3 μmol/L. Conclusion SeO2 can inhibit cell proliferation and up-regulate c-los expression in GLC-82 cells. This indicates that SeO2 may be involved in the process of apoptosis through c-los protein.