论文部分内容阅读
目的探讨蓬子菜黄酮类化合物香叶木素-7-O-β-D-木糖-(1→6)-β-D-葡萄糖苷(DXG)对人肝癌细胞株HepG2增殖的抑制作用及其诱导HepG2细胞凋亡的机制。方法台盼蓝染色法检测DXG对HepG2细胞生长的影响;MTT法检测DXG对细胞增殖的抑制作用;吖啶橙/溴乙(AO-EB)荧光染色法观察DXG对HepG2细胞凋亡的形态学影响;琼脂糖凝胶电泳观察凋亡细胞的DNA图谱变化;反转录聚合酶链式反应(RT-PCR)检测凋亡相关基因bcl-2及bax的表达。结果台盼蓝染色和MTT检测显示,DXG 50、100、200μg/mL能明显抑制HepG2细胞增殖,与对照组相比差异显著(P<0.05)。AO-EB染色可见DXG诱导HepG2细胞凋亡并发生形态学改变,且随DXG的质量浓度升高,凋亡细胞的比例显著增加。琼脂糖凝胶电泳显示,HepG2细胞经DXG 100、200μg/mL处理48 h后,可见DNA“梯形”碎片条带。RT-PCR实验表明,DXG下调细胞凋亡的抑制基因bcl-2 mRNA表达,促使凋亡基因bax mRNA表达增强。结论 DXG具有显著抑制人肝癌细胞HepG2增殖、诱导其凋亡的作用,其作用机制与调控bax/bcl-2的表达有关。
OBJECTIVE To investigate the inhibitory effect of flavonoid xanthophyllin -7-O-β-D-xylose- (1 → 6) -β-D-glucoside (DXG) on the proliferation of human hepatocellular carcinoma cell line HepG2 and its Mechanism of inducing HepG2 cell apoptosis. Methods The effect of DXG on the proliferation of HepG2 cells was detected by trypan blue staining. The inhibitory effect of DXG on the proliferation of HepG2 cells was detected by MTT assay. The apoptosis of HepG2 cells was observed by AO-EB staining. The changes of DNA profiles of apoptotic cells were observed by agarose gel electrophoresis. The expressions of bcl-2 and bax were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results Trypan blue staining and MTT assay showed that DXG 50, 100 and 200 μg / mL significantly inhibited the proliferation of HepG2 cells compared with the control group (P <0.05). AO-EB staining showed that DXG induced HepG2 cells apoptosis and morphological changes, and with the DXG concentration increased, the proportion of apoptotic cells increased significantly. Agarose gel electrophoresis showed that after HepG2 cells were treated with DXG 100 and 200 μg / mL for 48 h, DNA “trapezoid” fragments could be seen. RT-PCR experiments showed that DXG down-regulated the expression of bcl-2 mRNA, and promoted the expression of bax mRNA. Conclusion DXG can significantly inhibit the proliferation and induce apoptosis of human hepatocellular carcinoma HepG2 cells. Its mechanism may be related to the regulation of bax / bcl-2 expression.