黄连素抑制人喉癌 Hep-2细胞增殖及机制研究

目的：探讨黄连素对人喉癌 Hep-2细胞增殖的抑制作用及其可能机制。方法体外培养 Hep-2细胞，使用不同浓度黄连素诱导不同时间，应用 MTT 法检测细胞增殖抑制率；流式细胞术测定细胞周期及凋亡；实时荧光定量 PCR 和蛋白质免疫印迹（Western blot）法分别检测细胞周期蛋白 D（CyclinD）、B 细胞淋巴瘤/白血病-2（Bcl-2）、Bcl-2相关 X 蛋白（Bax）基因和蛋白表达水平。结果与0μmol/ L 黄连素对照组比较，2.5、5.0、10.0、20.0μmol/ L 黄连素作用 Hep-2细胞24、48 h，可明显抑制细胞增殖，且呈浓度时间依赖性（P ＜0.05）；2.5、5.0、10.0μmol/ L 黄连素作用 Hep-2细胞48 h，细胞早期凋亡率明显升高，呈浓度依赖性（P ＜0.05）；G0/ G1期细胞增多（P ＜0.05），诱导细胞阻滞于 G0/ G1期；10μmol/ L 黄连素作用 Hep-2细胞48 h，Bax 基因和蛋白表达水平明显升高（P ＜0.05），CyclinD、Bcl-2基因和蛋白表达水平明显下降（P ＜0.05）。结论黄连素能抑制喉癌细胞增殖，引起 G0/ G1期阻滞，并诱导其凋亡，其作用机制可能与 Bax 基因表达上调及 CyclinD、Bcl-2基因表达下调有关。

Berberine on proliferation of human laryngeal Hep-2 carcinoma cells

objective To investigate the effect of berberine on proliferation of human laryngeal Hep-2 carcinoma cells and its possible mechanism. Methods The Hep-2 cells were treated with different concentrations of berberine for dif-ferent hours,the proliferation inhibitory rates were detected by MTT assay;the cell cycle and apoptosis were analyzed by flow cytometry;the levels of CyclinD,Bcl-2 and Bax were measured by real-time PCR and Western blot. Results The proliferation inhibition rates of Hep-2 cells treated with 2. 5、5. 0、10. 0、20. 0 μmol/ L berberine for 24 and 48 hours were increased compared with 0μmol/ L group in concentration-and time-dependent manners(P < 0. 05). The early apoptotic rate was increased in concentration-dependent manner(P < 0. 05),and the proportion of cells in G0 /G1 phase was increased after treated with 2. 5,5. 0,10. 0 μmol/ L berberine for 48 hours(P < 0. 05). The levels of Bax mRNA and protein expression significantly increased(P < 0. 05),while the levels of CyclinD and Bcl-2 expres-sion significantly decreased(P < 0. 05). Conclusion Berberine can inhibit proliferation,and induct cell cycle G0 /G1 arrest and apoptosis of Hep-2 cells probably via upregulation of Bax and downregulation of CyclinD and Bcl-2 mR-NA and protein expression.