摘要
目的探讨淫羊藿苷对人小细胞肺癌细胞株NCI-H446的作用及其机制。方法将对数生长期NCI-H446细胞分成对照组和淫羊藿苷组。对照组细胞常规培养,淫羊藿苷组细胞培养体系中加入淫羊藿苷(8μmol/L)。各组细胞培养48 h后,利用噻唑蓝检测细胞增殖;流式细胞术检测细胞凋亡;实时定量聚合酶链式反应检测细胞Janus激酶2(JAK2)和信号转导子与转录激活子3(STAT3)基因表达;蛋白质印迹法检测细胞中JAK2、STAT3、磷酸化JAK2(p-JAK2)、磷酸化STAT3(p-STAT3)、Bax和BCL-2蛋白的变化。结果与对照组相比,淫羊藿苷组NCI-H446细胞增殖率明显下降(P<0.05),凋亡率明显升高,NCI-H446细胞JAK2 m RNA与STAT3 m RNA无明显差异。蛋白质印迹法检测显示,淫羊藿苷组NCI-H446细胞JAK2、STAT3总蛋白无明显变化(P>0.05);但p-JAK2、p-STAT3和Bax蛋白明显增加,而BCL-2明显减少(P<0.05)。结论淫羊藿苷能抑制NCI-H446细胞增殖,促进NCI-H446细胞凋亡,其作用可能是通过影响JAK2/STAT3信号传导通路实现的。
Objective To investigate the effect of icaritin on the small cell lung cancer cell lines NCI-H446 and its mechanism. Methods The NCI-H446 cells at logarithmic growth phase were divided into control and icaritin groups. The cells in the control group were normally treated and cells in the icaritin group were incubated with icaritin （8 μmol/L）. Thiazole blue and flow cytometry were used to examine the proliferation and apoptotic changes in the two groups 48 hours after incubation respectively. Gene expression of Janus kinase 2 （JAK2） and signal transducers and activators of transcription 3 （STAT3） were detected by real-time quantitative polymerase chain reaction. The changes of JAK2, STAT3, phospho-JAK2 （p-JAK2）, phospho-STAT3 （p-STAT3）, Bax and BCL-2 protein were detected by Western blotting. Results Compared with the control group, the proliferation rate of NCI-H446 cells in the icaritin group was significantly lower （P 〈 0.05）, but the apoptotic rate of NCI-H446 cells in the icaritin group was significantly higher （P 〈 0.05）. After the treatment with icaritin, the expression of JAK2 and STAT3 mRNA had no obvious differences. The Western blotting results showed that there was no significant changes in total JAK2, STAT3 protein （P 〉 0.05）, but an increasing trend in p-JAK2, p-STAT3 and Bax was observed with the decreasing of BCL-2 （P 〈 0.05）. Conclusion Icaritin can inhibit the proliferation and promote the apoptosis of NCI-H446 cells and the effect may be achieved through JAK2/STAT3 signal transduction pathway.
出处
《华西医学》
CAS
2015年第8期1401-1405,共5页
West China Medical Journal
