2,3-吲哚醌诱导人神经母细胞瘤细胞凋亡作用及机制

ISA INDUCES APOPTOSIS OF HUMAN NEUROBLASTOMA CELL LINE SH-SY5Y

目的观察2,3-吲哚醌(ISA)经Caspase-3途径诱导人神经母细胞瘤凋亡的作用及机制。方法以不同浓度ISA(0、100、200、400μmol/L)作用于SH-SY5Y细胞48 h,采用Hocehst33258/PI荧光染色技术观察细胞凋亡的形态学改变,流式细胞术检测细胞凋亡率的变化,Western blot方法检测Caspase激活的脱氧核糖核酸酶抑制剂(ICAD)蛋白表达的变化。结果Hochest33258/PI荧光染色可见核浓缩及核碎裂等典型的细胞凋亡形态学特征。100、200、400μmol/L ISA处理组SH-SY5Y细胞的凋亡率均明显高于对照组(F=111.30,P<0.05)。随着ISA浓度的升高,表达活化Caspase-3的细胞数逐渐增加(F=5.622,P<0.05)。Western blot检测到其下游作用底物ICAD被降解。结论ISA能诱导SH-SY5Y细胞凋亡,该作用可能通过激活Caspase-3来实现。

Objective To explore the effects and mechanisim of caspase-3 activation in ISA-induced apoptosis of human neurohlastoma cell line SH-SY5Y through caspase-3 pathway. Methods The apoptosis was detected by flow cytometry （FCM） and Hochest33258/PI staining, and the activity of caspase-3 determined by FCM. Western blot was used to analyze inhibitor of caspase-activated DNAse （ICAD）, the suhstrate of Caspase-3 after treatment of ISA at different concentrations （0, 100, 200, 400μmol/L）. Results The morphological features of nuclear condensation and fragmentation were observed after exposure to 400 μmol/L of ISA. The apoptotic rate of SH SY5Y cells, after treatment with ISA （100, 200, 400 μmol/L）, was significantly higher than that in the control. The activation of caspase-3 increased with an elevation of ISA density （F=5. 622,P〈0.05）. In hihitor of caspase-activated DNase （ICAD） protein, the suhstrate of Caspase-3, was degraded. Conclusion Isatin could induce apoptosis of SH-SY5Y cells, which, probably, is completed through the activation of caspase-3.