人参皂苷Rg3诱导结直肠癌细胞凋亡作用研究

Effect of Ginsenoside Rg3 on Apoptosis of Colorectal Cancer Cells

本文初步探讨人参皂苷Rg3(Rg3)体外抗结直肠癌作用研究,采用高效液相色谱法(High Performance Liquid Chromatography,HPLC)测定Rg3的含量,以流式细胞术法测定Rg3的凋亡百分数,并利用蛋白质印迹法(Western Blot,WB)测定c-Jun氨基端激酶(c-Jun N-terminal kinase,JNK)、磷酸化c-Jun氨基端激酶(Phosphorylation c-Jun N-terminal kinase,P-JNK),BCL2-Associated X的蛋白质(BAX)、B细胞淋巴瘤-2基因(B-cell lymphoma-2,BCL-2)和天冬氨酸特异性半胱氨酸蛋白酶3(cysteine-containing aspartate specific proteinase 3,Caspase 3)凋亡蛋白的表达情况。结果显示,Rg3出峰时间在8 min左右,纯度为≥98%;Rg3孵育细胞6 h、12 h和24 h后,细胞凋亡率分别为18.96%、31.90%和81.2%;BAX(21 kDa)在结直肠癌(colorectal cancer,CRC)细胞系中表达显著增加(P<0.01,P<0.01),而BCL-2(26 k Da)呈现完全相反的趋势(P<0.001,P<0.01),JNK激酶可激活内质网基膜上的促凋亡因子BAX构象改变而活化,并弱化BCL-2的抗凋亡能力。结果表明,Rg3具有很好的抗结直肠癌效果。

The resistance of ginsenoside Rg3(Rg3)against the colorectal cancer(CRC)in vitro was preliminarily investigated.Rg3 concentration was measured by the high-performance liquid chromatography mothod,and the apoptotic percentage of Rg3 was detected using the flow cytometry.The expressions of apoptosis proteins were monitored by Western blotting,including c-Jun N-terminal kinase(JNK),phosphorylated JNK(P-JNK),BCL2-Associated X protein(BAX),B-cell lymphoma-2(BCL-2),and cysteine-containing aspartate specific proteinase 3(Caspase 3).The retention time of Rg3 is about 8 min,and its purity is≥98%.After incubation with Rg3 for 6 h,12 h and 24 h,the cell apoptosis rates are 18.96%,31.90%and 81.2%,respectively.The BAX(21 k Da)is upregulated and Bcl-2(26 kDa)is downregulated significantly in CRC cell lines.JNK can promote the conformation variation and activation of the proapoptosis factor BAX on the endoplasmic reticulum basal membrane and weaken the anti-apoptotic ability of BCL-2.These findings show Rg3 has an excellent anti-CRC effect.