摘要
目的观察紫杉醇对人胆管癌RBE细胞凋亡及细胞外信号调节激酶 (ERK)表达的影响 ,以了解ERK在其中的作用。方法运用体外细胞培养 ,AnnexinV 异硫氰酸荧光素和碘化丙啶双染法及流式细胞技术观察紫杉醇以及与ERK上游激酶MEK抑制剂PD980 5 9合用 2 4h后RBE细胞的凋亡情况 ,运用蛋白质印迹法观察紫杉醇以及与PD980 5 9合用后RBE细胞的ERK1,ERK2和ERK的活化形式 (pERK)的表达情况。 结果紫杉醇诱导RBE细胞 ,凋亡率为 14 8%± 2 7% (n =6 ) ,明显高于对照组凋亡率 3 4 %± 1 3% (n =6 ) ,(P <0 0 1) ,紫杉醇合用PD980 5 9后凋亡率2 8 2 %± 5 7% (n =6 ) ,明显高于单用紫杉醇组凋亡率 14 8%± 2 7% (n =6 ) ,(P <0 0 1) ,紫杉醇可介导细胞 pERK的表达增加 ,两药合用后细胞的ERK1,ERK2及 pERK的表达降低。 结论紫杉醇可诱导胆管癌细胞凋亡 ,并激活了ERK途径 ,针对ERK的抑制剂可抑制紫杉醇介导的ERK的活化 ,同时增强紫杉醇诱导的凋亡作用。
Objective To investigate the effect on apoptosis and extracellular signal regulated kinase (ERK) expression of cholangiocarcinoma RBE cell line induced by paclitaxel and the role of extracellular signal regulated kinase in paclitaxel-induced apoptosis. Methods By techniques of cell culture in vitro, annexin-V-FITC and propidium iodide(PI)double staining and flow-cytometry (FCM), RBE cell line was treated by paclitaxel or together with MEK inhibitor PD98059 for 24 h. Using Western blot test the expression of ERK1, ERK2, pERK after RBE cell line was treated by paclitaxel or together with PD98059. Results Paclitaxel induced RBE cell line to apoptosis, the ratio of apoptosis was 14.8%±2.7%(n=6), the control apoptosis was 3.4%±1.3%(n=6,P<0.01). A combination of paclitaxel and PD98059 can enhance the apoptosis of RBE cell line and the ratio was 28.2%±5.7%(n=6, P<0.01). The level of pERK was significantly enhanced by treatment with paclitaxel and the combination of paclitaxel and PD98059, which resulted in down-regulation of ERK1, ERK2 and pERK. Conclusions Paclitaxel induced RBE cell line to apoptosis and activated ERK pathway. PD98059 blocked ERK activation by paclitaxel and enhanced apoptosis of RBE cell line induced by paclitaxel.
出处
《中华普通外科杂志》
CSCD
北大核心
2003年第11期684-686,共3页
Chinese Journal of General Surgery
