Stellettin B对非小细胞肺癌细胞增殖与凋亡的影响

目的 探讨Stellettin B 对非小细胞肺癌细胞增殖和凋亡的影响及可能的作用机制。方法采用四甲基偶氮唑盐（MTT）比色法检测不同浓度（01、1、10、100μmol／L）Stellettin B处理A549细胞和NCI-H1299细胞24、48、72和92h的增殖抑制率；采用Hoechst染色检测处理A549细胞24、48h后的凋亡指数；流式细胞术Annexin-FITC／PI双染法和PI染色法分别检测处理A549细胞48h后的凋亡率和细胞周期分布情况；采用Western blotting检测处理48h后A549细胞周期相关蛋白（Cyclin A、CDK2及p21CIP1）的表达水平。结果 不同浓度Stellettin B可显著增强对A549细胞的增殖抑制作用，且呈剂量和时间依赖性（P＜0.05）。不同浓度Stellettin B处理24、48h后，A549细胞的凋亡指数、凋亡率均升高且呈剂量依赖性（P＜0.05）。随着Stellettin B浓度的增加，G0／G1期细胞比例及p21CIP1蛋白的表达水平逐渐升高，S和G2／M期细胞比例及Cyclin A和CDK2蛋白的表达水平逐渐降低，差异均有统计学意义（P＜0.05）。结论 Stellettin B 可抑制A549细胞增殖并诱导细胞凋亡及细胞周期阻滞，可能与其对细胞周期相关蛋白表达的调控有关。

Effects of stellettin B on proliferation and apoptosis of non-small cell lung cancer cells

Objective To explore the effects of stellettin B on proliferation, apoptosis of non-small cell lung cancer（NSCLC） cells and its possible active mechanism. Methods The A549 and NCI-H1299 cells were treated with different concentrations of stelle- ttin B （0. 1, 1, 10, 100μmol/L）. The MTT method was used to measure the proliferation inhibition rate at 24^th, 48^th, 72^nd and 96^thh treated with different concentrations of stellettin B. The Hoechst staining was employed to detect the cell apoptosis index at 24th and 48^th h after treatment with stellettin B. The Annexin-FITC/PI double staining and PI staining were employed to detect cell apoptosis and cell cycle distribution at 48^th h via flow cytometry. The Western blotting was used to measure the protein levels of cell cycle-related genes （ eyclin A, CDK2 and p21^CIPI ） at 48^th h after treatment. Results The stellettin B of different concentrations could increase the prolif- eration inhibition rates in a dose and time-dependent manner. After being treated by stellettin B for 24 and 48h, there were increased apoptosis index, apoptosis rate in a dose-dependent manner. With the increase of stellettin B, there were increased G0/G1 phase cell percentage and protein level of p21^GIPI but decreased S and GJM phase cell percentages and protein level of eyclin A and CDK2 with statistical difference （P〈0. 05）. Conclusion Stellettin B can inhibit the proliferation of NSCLC cells as well as inducement of apopto- sis and cell cycle arrest, possibly by influencing the expressions of cycle-related genes.