黄芪多糖通过ANXA3诱导人结肠癌HT-29细胞凋亡

Astragalus polysaccharide induces the apoptosis of human colon cancer HT-29 cells through ANXA3

目的探讨黄芪多糖(RSP)对人结肠癌HT-29细胞增殖、凋亡的影响,并初步观察其作用机制。方法采用不同浓度(0.5、1.0、2.0、4.0 mg/mL)RSP、HY-N0168(2.0μg/mL)和MPT0B392(0.5μg/mL)处理HT-29细胞;CCK-8法检测RSP对HT-29细胞活力影响;流式细胞术分析HT-29细胞凋亡情况;Western blot方法分析RSP对HT-29细胞中total-PARP、Cle-PARP、Cle-caspase 3、ANXA3表达及p38和JNK磷酸化水平的影响。结果RSP呈浓度依赖性的抑制HT-29细胞活力、细胞周期进展及增殖;RSP还诱导HT-29细胞中Caspase-3/-9活性上调及细胞凋亡;RSP抑制HT-29细胞中ANXA3/p38和JNK信号通路活化;然而,p38激动剂HY-N0168和JNK激动剂MPT0B392能部分逆转RSP对HT-29细胞的毒性作用;RSP通过ANXA3降低p38和JNK信号通路活化及RSP细胞的毒性作用。结论RSP可通过降低ANXA3/p38和JNK信号通路转导抑制人结肠癌细胞增殖。

Objective To explore the effects of astragalus polysaccharide(RSP)on the proliferation and apoptosis of human colon cancer HT-29 cells,and further observe its mechanisms.Methods After treatment with RSP(0.5,1.0,2.0,4.0 mg/ml),HY-N0168(2.0μg/ml)and MPT0B392(0.5μg/ml),the effects of RSP on the cell viability were measured by using CCK-8 assay.The cell apoptosis was detected by flow cytometry.Western blot assay was used to investigate the impacts of RSP on the expression levels of total-PARP,Cle-PARP,Cle-caspase 3,ANXA3 p-p38 and p-JNK in HT-29 cells.Results RSP dose-dependently inhibited survival,cell cycle progression and proliferation of HT-29 cells.RSP induced caspase-3/-9 and apoptosis activation in HT-29 cells.RSP suppressed the activation of ANXA3/p38 and JNK pathways in HT-29 cells.Conversely,its cytotoxicity against HT-29 cells was largely attenuated by co-treatment of p38 activator HY-N0168 and the JNK activator MPT0B392.RSP decreased the activation of p38 and JNK cascades and its cytotoxicity against HT-29 cells through ANXA3.Conclusion RSP could inhibit the proliferation of human colon cancer cells via decreasing the activation of ANXA3/p38 and JNK cascades.