硝酸甘油对肿瘤细胞增殖及蛋白表达的调控

Regulation of proliferation and protein expression in GTN treated tumor cells

目的探讨硝酸甘油(GTN)作为一氧化氮供体对人宫颈癌Hela细胞增殖及全细胞蛋白质组学表达的调控意义。方法分别用细胞形态学和噻唑蓝比色法(MTT)观察经GTN作用的Hela细胞生长和增殖状况;双向凝胶电泳技术分离Hela细胞蛋白质,观察GTN对细胞蛋白表达谱的影响。结果培养液中GTN浓度为40μg/ml时Hela细胞分离、皱缩,出现小球状或异型性,细胞外杂质增多,密度明显下降(0.0825,P<0.05);MTT检测细胞增殖抑制率为73.3%;双向电泳分析显示GTN组与对照组Hela细胞蛋白质点匹配率为86%,GTN组较对照组312个(47%)蛋白点表达明显改变,101个(15.2%)上调,211个(31.8%)下调,且有17个蛋白点只在GTN组细胞中表达,21个蛋白点只在对照组细胞中表达。结论一定浓度的GTN能够抑制肿瘤细胞的生长和调控细胞内蛋白质表达。

Objective To investigate the proliferation and analyze the protein expression of the Hela cells by the regulation of glyceryl trinitrate （GTN） that acts as the donor of nitric oxide. Methods Cell morphology and MTT assay were used to analyze cell proliferation. Two-dimensional polyacrylamide gel electrophoresis （2-DE） and computer-assisted image analysis find the spots of interests. Difference of protein expression between GTN-treating group and control group was detected. Results Hela cells treated with 40μg/ml GTN in culture medium underwent inhibition of the growth activity, which were not clustered and shriveled. Some atypia cells were observed. Density of the cells was obviously decreased （0. 0825, P 〈 0. 05）. The inhibition ratio of cell proliferation was 73. 3% in MTT assay. The matching rate of the GTN group was 86%. Compared with control group, there were 312 protein spots changed greatly in GTN-treating group, which had 101 （ 15.2% ） protein spots up-regulated and 211 （31.8%） protein spots down-regulated. 17 proteins were detected only in GTN-treating group cells and 21 proteins were detected only in control group cells. Conclusion Glyceryl trinitrate in certain concentration could inhibit the Hela cells in vitro and the spots of interests may be identified as the proteins that associate with regulation of GTN, which will be further analyzed.