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目的:研究紫草素诱导人前列腺癌细胞(PC-3)凋亡及其作用机制。方法:以PC-3细胞为研究对象,以不同浓度的紫草素处理PC-3细胞,另设空白组,处理不同时间后,分别采用噻唑蓝(MTT)法检测紫草素对PC-3细胞增殖抑制情况;显微镜观察细胞形态变化;流式细胞仪检测细胞中活性氧(reactive oxygen species,ROS)的产生;蛋白质免疫印迹(Western blot)检测B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax),半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)和磷酸化-细胞外信号调节激酶1/2(p-ERK1/2)蛋白的表达。结果:不同浓度的紫草素对PC-3细胞的生长抑制呈时间和剂量依赖性;形态学观察结果显示紫草素作用PC-3细胞后可使细胞变圆,脱落,皱缩,细胞内出现空泡,周围出现凋亡小体;与空白组比较,不同浓度的紫草素明显升高Caspase-3和Bax蛋白的表达,明显降低Bcl-2蛋白的表达,不同浓度的紫草素作用细胞48 h后可诱导细胞产生ROS,ROS清除剂二硫苏糖醇(DTT)可以明显逆转紫草素诱导的PC-3细胞的生长抑制率,且0.2 mmol·L-1DTT可以抑制由紫草素诱导的Caspase-3和ERK1/2的活化。结论:紫草素通过ROS/ERK1/2信号通路诱导人前列腺癌PC-3细胞发生凋亡。
Objective: To study the apoptosis induced by shikonin in human prostate cancer cells (PC-3) and its mechanism. Methods: PC-3 cells were treated with different concentrations of shikonin and PC-3 cells were treated with different concentrations of shikonin. The cells were treated with different concentrations of shikonin (MTT) The changes of cell morphology were observed under microscope. The production of reactive oxygen species (ROS) was detected by flow cytometry. The expressions of Bcl-2 and Bcl-2 were detected by Western blot. ), Expression of Bcl-2, caspase-3 and p-ERK1 / 2 . RESULTS: Different concentrations of shikonin inhibited the growth of PC-3 cells in a time-and dose-dependent manner. Morphological observation showed that shikonin could cause rounding, shedding and shrinking of cells in PC-3 cells, Compared with the blank group, different concentrations of shikonin significantly increased the expression of Caspase-3 and Bax protein, significantly decreased the expression of Bcl-2 protein, different concentrations of shikonin After 48 h, cells were induced to produce ROS. ROS scavenger dithiothreitol (DTT) could obviously reverse the growth inhibition rate of PC-3 cells induced by shikonin, and 0.2 mmol·L-1 DTT could inhibit the proliferation of PC Superoxide-induced activation of Caspase-3 and ERK1 / 2. Conclusion: Shikonin induces apoptosis in human prostate cancer PC-3 cells through ROS / ERK1 / 2 signaling pathway.