Jiang Shu-Xian, Qi Bo, Yao Wen-Jian, Gu Cheng-Wei, Wei Xiu-Feng, Zhao Yi, Liu Yu-Zhen, Zhao Bao-Sheng
Shu-Xian Jiang, Bo Qi, Wen-Jian Yao, Cheng-Wei Gu, Xiu-Feng Wei, Yi Zhao, Yu-Zhen Liu, Bao-Sheng Zhao, Department of Thoracic Surgery, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, Henan Province, China.
World J Gastroenterol. 2017 Apr 14;23(14):2511-2518. doi: 10.3748/wjg.v23.i14.2511.
To investigate the effects of berberine on esophageal cancer (EC) cells and its molecular mechanisms.
Human esophageal squamous cell carcinoma cell line KYSE-70 and esophageal adenocarcinoma cell line SKGT4 were used. The effects of berberine on cell proliferation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For cell cycle progression, KYSE-70 cells were stained with propidium iodide (PI) staining buffer (10 mg/mL PI and 100 mg/mL RNase A) for 30 min and cell cycle was analyzed using a BD FACSCalibur flow cytometer. For apoptosis assay, cells were stained with an Annexin V-FITC/PI apoptosis detection kit. The rate of apoptotic cells was analyzed using a dual laser flow cytometer and estimated using BD ModFit software. Levels of proteins related to cell cycle and apoptosis were examined by western blotting.
Berberine treatment resulted in growth inhibition of KYSE-70 and SKGT4 cells in a dose-dependent and time-dependent manner. KYSE-70 cells were more susceptible to the inhibitory activities of berberine than SKGT4 cells were. In KYSE-70 cells treated with 50 μmol/L berberine for 48 h, the number of cells in G/M phase (25.94% ± 5.01%) was significantly higher than that in the control group (9.77% ± 1.28%, < 0.01), and berberine treatment resulted in p21 up-regulation in KYSE-70 cells. Flow cytometric analyses showed that berberine significantly augmented the KYSE-70 apoptotic population at 12 and 24 h post-treatment, when compared with control cells (0.83% 43.78% at 12 h, < 0.05; 0.15% 81.86% at 24 h, < 0.01), and berberine-induced apoptotic effect was stronger at 24 h compared with 12 h. Western blotting showed that berberine inhibited the phosphorylation of Akt, mammalian target of rapamycin and p70S6K, and enhanced AMP-activated protein kinase phosphorylation in a sustained manner.
Berberine is an inhibitor of human EC cell growth and could be considered as a potential drug for the treatment of EC patients.
研究小檗碱对食管癌（EC）细胞的作用及其分子机制。
使用人食管鳞状细胞癌细胞系KYSE - 70和食管腺癌细胞系SKGT4。采用3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐（MTT）法评估小檗碱对细胞增殖的影响。对于细胞周期进程，用碘化丙啶（PI）染色缓冲液（10 mg/mL PI和100 mg/mL核糖核酸酶A）对KYSE - 70细胞染色30分钟，然后使用BD FACSCalibur流式细胞仪分析细胞周期。对于凋亡检测，用膜联蛋白V - FITC/PI凋亡检测试剂盒对细胞进行染色。使用双激光流式细胞仪分析凋亡细胞率，并使用BD ModFit软件进行估算。通过蛋白质印迹法检测与细胞周期和凋亡相关的蛋白质水平。
小檗碱处理以剂量和时间依赖性方式导致KYSE - 70和SKGT4细胞生长抑制。KYSE - 70细胞比SKGT4细胞对小檗碱的抑制活性更敏感。在用50 μmol/L小檗碱处理48小时的KYSE - 70细胞中，G/M期细胞数量（25.94% ± 5.01%）显著高于对照组（9.77% ± 1.28%，P < 0.01），且小檗碱处理导致KYSE - 70细胞中p21上调。流式细胞术分析表明，与对照细胞相比，小檗碱在处理后12小时和24小时显著增加了KYSE - 70凋亡细胞群体（12小时时从0.83%增加到43.78%，P < 0.05；24小时时从0.15%增加到81.86%，P < 0.01），且小檗碱诱导的凋亡效应在24小时比12小时更强。蛋白质印迹法显示，小檗碱持续抑制Akt、雷帕霉素靶蛋白和p70S6K的磷酸化，并增强AMP激活的蛋白激酶磷酸化。
小檗碱是人类EC细胞生长的抑制剂，可被视为治疗EC患者的潜在药物。
