小檗碱抗人结直肠腺癌的作用及其机制

Anticancer property of berberine in human colorectal adenocarcinoma

目的探讨黄连的主要药用成分小檗碱的抗结直肠腺癌的机制。方法不同浓度的小檗碱作用于人结直肠腺癌(HCT-15)细胞后,用显微镜观察人结直肠腺癌细胞的形态学改变;细胞染色方法观察细胞凋亡和自噬;细胞计数分析(CCK-8)检测小檗碱对细胞增殖的影响;免疫印迹法检测小檗碱对人结直肠癌细胞自噬和凋亡相关指标的影响;裸鼠移植瘤模型观察小檗碱的体内抑制肿瘤生长活性。结果显微镜观察结果显示小檗碱作用HCT-15细胞后细胞数量减少、间距增大、皱缩。小檗碱能够抑制HCT-15细胞增殖,对HCT-15细胞增殖的半数抑制浓度(IC_(50))为50μmol/L。免疫荧光和印迹法显示0、50、100μmol/L的小檗碱处理细胞不同时间后,细胞凋亡和自噬对应的标志物明显上升。免疫印迹结果显示小檗碱诱导HCT-15细胞自噬和凋亡从而抑制细胞的增殖。小檗碱诱导细胞自噬和凋亡主要通过激活p38途径。裸鼠实验结果表明小檗碱能够有效缩小体内肿瘤体积,0、10、20 mg/kg的小檗碱的抑瘤率分别为0、32%、74%。结论小檗碱在体外对HCT-15细胞增殖具有明显抑制作用,其机制主要是通过激活p38信号通路促进肿瘤细胞自噬和凋亡,并能有效抑制体内肿瘤的生长。

Objective To illustrate the anticancer property induced by berberine, which is extracted from Chinese medicine CoptidisRhizoma. Methods Human colorectal adenocarcinoma cells, HCT-15, were treated with different concentration of berberine. Cells were then observed under the optical microscope to analyze the morphological changes; CCK-8 assay was used to detect the inhibition of cell proliferation of HCT-15; Immunoblotting was used to detect the changes of autophagy and apoptosis related markers. Athymic mice injected with HCT-15 cells were used to detect the pharmacological activity of berberine in vivo. Results After berberine treatment, with the increased dose of berberine, the cell number reduced, cell spacing increased, and cell shape shrunk. The IC_（50） for berberine treatment in HCT-15 cell was 50 μmol/L. Autophagy and apoptosis markers were enhanced after treatment of 0, 50, and 100 μmol/L of berberine. The data further showed that enhanced autophagy and apoptosis were the main contributors for inhibiting cell proliferation. Immunoblotting analysis showed that berberine induced HCT-15 cells to autophagy and apoptosis mainly through p38 pathway. Athymic mice study showed that berberine treatment could decrease the volume of tumor in vivo, and the tumor inhibitory rates by berberine at 0, 50, and 100 mg/kg were 0, 32% and 74%, respectively. Conclusion Berberine inhibits the HCT-15 cell proliferation, mainly by p38 pathway and enhances autophagy and apoptosis. Berberine can effectively inhibit tumor growth in vivo.