吴茱萸碱调控人结肠癌HCT-116细胞中JAK2/STAT3信号通路的机制研究

摘要

Aim To investigate the inhibitory effect of Evodiamine on JAK2/STAT3 signal pathway in human colorectal cancer cell line HCT-116 . Methods Cells were cultured with 6. 0 μmol·L-1 Evodiamine for 2, 4 and 6 h, respectively. Cell nuclear morphology was detected by Hoechst staining and protein expression levels of JAK2 , p-JAK2 , STAT3 and p-STAT3 were examined by Western blot. Cells were treated with dif-ferent concentrations of AG490 for 48 h to select proper working concentration and cells treated with 6 μmol · L-1 EVO and 50 μmol · L-1 AG490 to compare the modulatory effect of EVO with AG490 on JAK2/STAT3 signal pathway. Results Hoechst staining revealed that Evodiamine could induce cells apoptosis, chroma-tin condensation gathered and typical apoptotic mor-phological changes in a time-dependent manner;West-ern Blot suggested that EVO could inhibit p-STAT3 significantly. After treatment with AG490, JAK2/STAT3 signal pathway was inactivated, the inhibitory effect of EVO on p-STAT3 was stronger than that of AG490 , while EVO combined with AG490 could fur-ther inhibit the expression of p-STAT3 significantly. Conclusions The anticancer effect of Evodiamine is mainly mediated by the modulation of JAK2/STAT3 signal pathway in HCT-116 cells.%目的：探讨吴茱萸碱( evodiamine, EVO)对人结肠癌HCT-116细胞中 JAK2/STAT3信号通路的影响。方法EVO诱导人结肠癌HCT-116细胞2、4、6 h后, Hoechst法检测细胞核凋亡的形态学改变；6μmol · L-1的 EVO 作用于HCT-116细胞2、4和6 h,不同浓度的AG490作用于 HCT-116细胞48 h,6μmol · L-1的 EVO 和50μmol · L-1的AG490分别诱导HCT-116细胞以及两药联合诱导HCT-116细胞6 h后,运用蛋白质印迹法检测各组细胞中 JAK2、p-JAK2、STAT3和 p-STAT3的蛋白表达量。结果镜下观察EVO诱导后的细胞核浓缩聚集,染色质边缘化,并出现染色质碎裂成块,形成凋亡小体等形态学改变。 Western blot结果显示：EVO可以明显下调HCT-116细胞内p-STAT3的表达；AG490可以抑制JAK2/STAT3信号通路的激活；EVO对JAK2/STAT3信号通路中 p-STAT3蛋白的表达抑制效果较AG490更明显,且两药联合应用后抑制效果进一步增强。结论 EVO对人结肠癌HCT-116细胞的抗肿瘤活性有可能是通过抑制JAK2/STAT3信号通路的激活来发挥的。