白皮杉醇调节JAK2/STAT3信号通路对胆囊癌细胞恶性生物学行为的影响

Influence of piceatannol on malignant biological behavior of gallbladder cancer cells by regulating JAK2/STAT3 signaling pathway

目的探讨白皮杉醇(PIC)基于酪氨酸激酶2/信号转导和转录激活因子3(JAK2/STAT3)信号通路对于胆囊癌细胞恶性生物学行为的影响。方法CCK-8法检测不同浓度PIC对胆囊癌GBC-SD细胞活性的影响。将细胞分为对照组(control组)、白皮杉醇低剂量组(PIC-L组,10μmol/L)、白皮杉醇中剂量组(PIC-M组,20μmol/L)、白皮杉醇高剂量组(PIC-H组,40μmol/L)、阳性对照组(吉西他滨,4 mg/L),结晶紫染色观察细胞平板克隆能力;划痕实验和Transwell实验检测GBC-SD细胞迁移和侵袭能力;流式细胞仪检测细胞凋亡;Western blot检测检测凋亡、迁移侵袭及JAK2/STAT3信号通路蛋白表达。结果选择PIC浓度10、20、40μmol/L进行后续实验。与control组比较,PIC-L、PIC-M、PIC-H组及吉西他滨组GBC-SD细胞的克隆形成数、迁移侵袭能力、MMP-2、MMP-9蛋白表达、p-JAK2/JAK2、p-STAT3/STAT3比值均显著下降(P<0.05);细胞凋亡率及凋亡蛋白(c-caspase-3、c-caspase-9)表达显著上升(P<0.05)。结论白皮杉醇可抑制胆囊癌GBC-SD细胞增殖、迁移和侵袭,诱导胆囊癌细胞GBC-SD凋亡,可能通过抑制JAK2/STAT3信号通路实现。

Objective To investigate the influence of piceatannol(PIC)on the malignant biological behavior of gallbladder cancer cells by regulating janus tyrosine kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway.Methods Cell Counting Kit-8(CCK-8)was used to detect the effect of different concentrations of PIC on human gallbladder carcinoma cells(GBC-SD)activity.The cells were divided into control group,low-dose PIC group(PIC-L,10μmol/L),medium-dose PIC group(PIC-M,20μmol/L),high-dose PIC group(PIC-H,40μmol/L)and positive control group(gemcitabine,4mg/L).Crystal violet staining was used to observe the cloning ability of cell plates.Scratch test and Transwell test were used to detect the migration and invasion abilities of GBC-SD cells.Cell apoptosis was detected by flow cytometry.Western blot was applied to detect the expression of apoptosis,migration and invasion,and JAK2/STAT3 signal pathway proteins.Results In this study,10,20 and 40μmol/L PIC was selected for subsequent experiments.Compared with the control group,the number of clone formation,migration and invasion abilities,the protein expression of matrix metalloproteinase 2(MMP-2)and matrix metalloproteinase 9(MMP-9),and the ratios of phosphorylated(p)-JAK2/JAK2 and p-STAT3/STAT3 decreased obviously in PIC-L,PIC-M,PIC-H and positive control groups(P<0.05).However,the apoptosis rate and the expression of apoptosis proteins(c-caspase-3 and c-caspase-9)increased significantly in PIC-L,PIC-M,PIC-H and positive control groups(P<0.05).Conclusion PIC can inhibit the proliferation,migration and invasion of GBC-SD cells and induce the apoptosis of GBC-SD cells possibly by inhibiting the JAK2/STAT3 signaling pathway.