莪术醇对乳腺癌细胞增殖凋亡及JAK2/STAT3信号通路的影响

Effect of Curcumol on JAK2/STAT3 Signaling Pathway in Breast Cancer Cells

[目的]探讨莪术醇对乳腺癌细胞增殖凋亡及JAK2/STAT3信号通路的影响。[方法]以不同浓度(0、12.5、25、50、100μg/ml)莪术醇作用于乳腺癌MCF-7细胞,MTT法检测MCF-7细胞增殖抑制率,流式细胞术观察细胞周期凋亡分布,Hoechst33258染色法观察细胞凋亡形态学变化,逆转录聚合酶链反应法检测细胞酪氨酸激酶2(JAK2)、信号转导子和转录激活子3(STAT3)信使核糖核酸相对表达量,蛋白印迹法检测细胞磷酸化JAK2(P-JAK2)、磷酸化STAT3(P-STAT3)蛋白相对表达量。[结果]莪术醇对MCF-7细胞增殖具有明显抑制作用,且抑制率随药物作用时间及作用浓度的增加而上升。随着莪术醇作用浓度的增加,细胞凋亡率也有所上升[(4.27%±0.82%)、(4.54%±1.21%)、(19.32%±1.87%)、(29.58%±2.92%)、(33.74%±3.91%)、(42.94%±2.81%)]。细胞周期结果显示,莪术醇可将细胞周期阻滞在G0/G1期,从而抑制细胞增殖。Western blot结果显示,莪术醇可抑制JAK2、STAT3磷酸化的表达,而RT-PCR结果显示,莪术醇作用细胞后,JAK2、STAT3 mRNA表达受到明显抑制。[结论]莪术醇作用于乳腺癌MCF-7细胞G0/G1期,从而抑制细胞增殖,诱导细胞凋亡,其机制可能是通过JAK2/STAT3信号通路来实现。

[Objective]To investigate the effect of curcumol on JAK2/STAT3 signaling pathway in breast cancer cells.[Methods]Breast cancer MCF-7 cells were treated with different concentrations of curcumol(0,12.5,25,50,100μg/ml).The proliferation inhibition rate of MCF-7 cells was detected by MTT assay.Cell apoptosis was observed by flow cytometry.Hoechst33258 staining method was used to observe the morphological changes of cell apoptosis.The relative expression of JAK2 and STAT3 m RNA was detected by RT-PCR.The relative expression of P-JAK2 and PSTAT3 protein was detected by Western blot.[Results]The curcumol had a significant inhibitory effect on the proliferation of MCF-7 cells,and the inhibition rate increased with the time and concentration of the drug.As the concentration of curcumol increased,the apoptosis rate also increased[(4.27%±0.82%),(4.54%±1.21%),(19.32%±1.87%),(29.58%±2.92%),(33.74%±3.91%),(42.94%±2.81%)].Cytometry showed that curcumol arrested the cell cycle in G0/G1 phase,thereby inhibiting cell proliferation.Western blot showed that curcumol inhibited the expression of JAK2 and STAT3 phosphorylation,and RT-PCR showed that JAK2 and STAT3 m RNA expression was significantly inhibited after cells were treatment with curcumol.[Conclusion]Curcumol acts on G0/G1 phase of breast cancer MCF-7 cells,inhibiting cell proliferation and inducing apoptosis,in which JAK2/STAT3 signaling pathway may be involved.