目的：探讨欧前胡素对人肝癌细胞的抑制作用及机制。方法将人肝癌细胞系HepG2用5、10、20、40、80μM的欧前胡素处理24h或48h，以相同培养条件不加欧前胡素的HepG2细胞为对照组，MTT法检测欧前胡素治疗后HepG2细胞的增殖抑制情况；HepG2细胞用10、20、40μM的欧前胡素处理24h，通过流式细胞术和Western blot检测欧前胡素对HepG2细胞的凋亡诱导作用和Mcl-1（myeloid cell leukemia-1）的表达；在HepG2细胞中转染pcDNA-3.1-Mcl-1真核表达载体后再用欧前胡素治疗，检测欧前胡素对HepG2细胞的增殖抑制作用和凋亡诱导效应。结果10、20、40、80μM欧前胡素组增殖抑制率分别为24h：（7.2±1.7）%、（14.5±2.6）%、（37.4±3.7）%、（56.3±4.3）%；48h：（14.5±2.1）%、（23.6±3.0）%、（52.3±4.8）%、（78.3±5.9）%，与对照组（0）比较，差异均有统计学意义（P<0.05）；10、20、40μM欧前胡素组凋亡诱导率分别为（3.9±0.8）%、（12.4±1.8）%、（26.9±2.5）%，与对照组（0.5±0.2）%比较，差异有统计学意义（P<0.05）。欧前胡素治疗后，HepG2细胞的Mcl-1表达水平显著下降，用pcDNA-3.1-Mcl-1真核表达载体转染HepG2细胞后，欧前胡素对HepG2细胞的凋亡诱导率较空pcDNA3.1载体转染组显著下降[（6.8±1.2）%比（25.7±2.3）%，P<0.05]。结论欧前胡素具有体外抗肝肿瘤的生物活性，其抗肿瘤效应的机制可能是通过下调Mcl-1蛋白表达从而诱导细胞凋亡。%Objective To investigate the inhibitory effect of imperatorin on human hepatocellular carcinoma cells and the underlying mechanism. Methods HepG2 cells were treated with 0 (control group), 5, 10, 20, 40, and 80μM imperatorin for 24 or 48 h, then MTT assay was used to detect the proliferation of HepG2 cells. The apoptosis of HepG2 cells and the expression of Mcl-1 was assessed by flow cytometry and western blot in HepG2 cells treated with 10, 20, 40 μM imperatorin for 24 h. The vector of pcDNA-3.1-Mcl-1 was transfected in HepG2 cells before treated with imperatorin, then the proliferation and apoptosis of cells were detected. Results The inhibition rate of proliferation of HepG2 cells treated with 10, 20, 40, and 80 μM imperatorin was as follows:(7.2±1.7)%, (14.5±2.6)%, (37.4±3.7)%, and (56.3±4.3)% for 24h;(14.5±2.1)%, (23.6±3.0)%, (52.3±4.8)%, and (78.3±5.9)% for 48h, all with significant difference compared with control group (P<0.05). The apoptosis rate of 10, 20, and 40μM group was (3.9±0.8)%, (12.4±1.8)%, and (26.9±2.5)%, respective, all with significant differences com-pared with that of control group[(0.5±0.2)%, P<0.05]. Imperatorin treatment significantly decreased the expression of Mcl-1. Transfection of pcDNA-3.1-Mcl-1 inhibited the anti-tumor effect of imperatorin [(6.8±1.2)% vs (25.7±2.3)%, P<0.05]. Conclusion Imperatorin exerts anti-tumor bio-activity. It may induce apoptosis of HepG2 cells via down-regulating the expression of Mcl-1.
展开▼
