藤黄酸通过抑制JAK-STAT通路对食管癌细胞KYSE450增殖和凋亡的影响

Eff ect of gambogic acid on proliferation and apoptosis of esophageal cancer cell KYSE450 by inhibiting JAK-STAT pathway

目的探讨藤黄酸对食管癌细胞KYSE450的增殖和凋亡以及对Janus激酶(Janus kinase,JAK)-信号转导与转录激活子(signal transducer and activator of transcription,STAT)信号通路的影响。方法常规培养食管鳞癌细胞KYSE450,分别加入0、0.5、1.0和2.0μmol/L浓度的藤黄酸处理24、48和72 h后,采用CCK-8实验检测细胞的存活率;观察药物处理细胞的状态和集落形成能力;transwell实验检测细胞的侵袭和迁移能力;流式细胞术检测细胞的凋亡率;Western blot实验检测细胞中JAK-STAT通路的相关蛋白[JAK2、STAT3、磷酸化的Janus激酶2(phosphorylated Janus kinase 2,P-JAK2)和P-STAT3]和凋亡相关蛋白(Bcl2、Bax、Cleaved PARP1、Cleaved Caspase-3和Cleaved Caspase-9)的表达情况。结果经过0、0.5、1.0和2.0μmol/L浓度的藤黄酸分别处理24、48和72 h后,食管癌KYSE450细胞增殖均被抑制,并呈时间和浓度依赖性(均P<0.05)。经0、0.5和1.0μmol/L藤黄酸处理24 h的食管癌细胞KYSE450的侵袭和迁移的能力减弱(均P<0.05)。0.5和1.0μmol/L的藤黄酸处理24 h的KYSE450细胞凋亡比例均增加(均P<0.05)。Western blot结果显示,0.5和1.0μmol/L藤黄酸处理48 h后,KYSE450细胞中Bax、Cleaved PARP 1、Cleaved Caspase-3和Cleaved Caspase-9蛋白表达量均增加,Bcl2、P-JAK2和P-STAT3蛋白表达量均降低(均P<0.05)。结论藤黄酸可以抑制食管癌细胞的增殖,并且通过JAK-STAT信号通路诱导食管癌细胞凋亡。

Objective To investigate the effect of gambogic acid on the proliferation and apoptosis of esophageal cancer cell KYSE450 and the Janus kinase(JAK)-signal transducer and activator of transcription(STAT)signaling pathway.Methods Esophageal cancer cell KYSE450 was routinely cultured and treated with gambogic acid at concentrations of 0,0.5,1.0 and 2.0μmol/L for 24,48,and 72 h,respectively.The survival rate of cells was measured by CCK-8 experiment.The status and colony formation ability of the drug-treated cells were observed.Transwell assay was used to detect the ability of cell invasion and migration.The apoptosis rate was detected by flow cytometry.Western blot test was used to detect JAK2,STAT3,phosphorylated Janus kinase 2(P-JAK2),P-STAT3 and apoptosis-related proteins(Bcl2,Bax,cleaved PARP1,cleaved caspase-3 and cleaved caspase-9).Results After treatment with 0,0.5,1.0,and 2.0μmol/L gambogic acid for 24,48,and 72 h,respectively,the proliferation of esophageal cancer cell KYSE450 was inhibited in a time-and concentration-dependent manner(all P<0.05).After treatment with 0,0.5 and 1.0μmol/L gambogic acid for 24 h,the invasion and migration ability of esophageal cancer cell KYSE450 was decreased (all P<0.05). The apoptosis ratios of KYSE450 cells treated with 0.5and 1.0 μmol/L gambogic acid for 24 h were increased (both P<0.05). Western blot results showed that the protein levels of Bax, cleavedPARP1, cleaved caspase-3 and cleaved caspase-9 were increased after treatment with 0.5 and 1.0 μmol/L gambogic acid for 48 h (allP<0.05), and the protein levels of Bcl2, P-JAK2 and P-STAT3 were decreased (all P<0.05). Conclusion Gambogic acid can inhibit theproliferation of esophageal cancer cells and induce the apoptosis of esophageal cancer cells through the JAK-STAT pathway.