重楼醇提取物通过JAK/STAT信号通路对神经胶质瘤U251细胞增殖及凋亡影响的研究

Effect of Rhizoma Paridis-alcohol Extract on Proliferation and Apoptosis of U251 Glioma Cells via JAK/STAT Signaling Pathway

目的:探讨重楼醇提取物通过酪氨酸激酶/信号转导子和转录激活子(JAK/STAT)信号通路对人神经胶质瘤U251细胞增殖及凋亡的影响。方法:用重楼醇提取物溶液干预人神经胶质瘤细胞U251细胞,然后采用MTT法检测重楼醇提取物对U251细胞生长增殖的影响。使用AnnexinⅤ-FITC/PI染色检测重楼醇提取物对U251细胞周期及凋亡的影响。Western blot及RT-PCR法检测重楼醇提取物对U251细胞磷酸化酪氨酸激酶2 (p-JAK2)、磷酸化信号传导及转录激活因子3 (p-STAT3)、细胞周期蛋白D1(Cyclin D1)、B淋巴细胞瘤-2基因(Bcl-2)蛋白及JAK2、STAT3、Cyclin D1、Bcl-2 mRNA表达的影响。结果:与空白组比较,重楼醇提取物各剂量组细胞抑制率、24 h后细胞凋亡率显著增加(P<0.05),且随着剂量的增加而增加(P<0.05,P<0.01);p-JAK2、p-STAT3、Cyclin D1及Bcl-2蛋白及JAK2、STAT3、Cyclin D1、Bcl-2 mRNA表达量均显著降低(P<0.05),且均随着剂量的增加而降低(P<0.05,P<0.01)。结论:重楼醇提取物可明显抑制人神经胶质瘤U251细胞的增殖,促进其凋亡,且其作用机制可能与重楼醇提取物阻滞JAK/STAT信号通路的信号传导,抑制通路蛋白及mRNA合成有关。

Objective: To investigate the effect of rhizoma paridis-alcohol extract on the proliferation and apoptosis of human U251 glioma cells via the signaling pathway of Janus kinase/signal transducer and activator of transcription(JAK/STAT).Methods: Treated U251 cells with rhizoma paridis-alcohol extract;detected its effect on the growth and proliferation of human U251 glioma cells by MTT assay,on the cell cycle and apoptosis by Annexin V-FITC/PI Apoptosis Detection Kit,and on the protein and mRNA expressions of phosphorylated Janus kinase-2(p-JAK2), phosphorylated signal transducer and activator of transcription-3(p-STAT3),Cyclin D1 and B-cell lymphoma-2(Bcl-2) by Western blot and RT-PCR. Results:The cell inhibition rate and apoptosis rate after 24 h in the rhizoma paridis-alcohol extract groups of different doses were increased significantly when compared with those in the blank group(P<0.05), and they were increased as the dose increased(P<0.05,P<0.01);the protein expression of p-JAK2,p-STAT3,Cyclin D1 and Bcl-2 and the mRNA expressions of JAK2、STAT3、Cyclin D1、Bcl-2 were significantly decreased(P<0.05),and they were decreased as the dose increased(P<0.05,P<0.01). Conclusion:The rhizoma paridis-alcohol extract can significantly inhibit the proliferation of human U251 glioma cells and promote its apoptosis, whose mechanism may be related to its blocking of the signal transduction of the JAK/STAT signaling pathway and inhibition of the synthesis of pathway proteins and mRNA.