酪氨酸激酶抑制剂对人囊肿衬里上皮细胞增殖及信号转导通路的影响  

Effects of tyrosine kinase inhibitor CL-387,785 on the proliferation and signal transduction pathway in human autosomal dominant polycystic kidney disease cyst-lining epithelial cells

作　　者：李林[1] 孙田美[1] 梅长林[1] 徐成钢[1] 赵海丹[1] 张树忠[1] 周玉坤[1] 

机构地区：[1]第二军医大学长征医院肾内科 解放军肾脏病中心,上海200003

出　　处：《中华肾脏病杂志》2003年第6期339-343,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金(30170901);国家科技部重大科技专项基金(2002AA2Z3130);上海市科委重大基础项目基金(02JC14029)

摘　　要：目的 研究表皮生长因子受体(EGFR)酪氨酸激酶抑制剂CL-387,785对人常染色体显性多囊肾病(ADPKD)囊肿衬里上皮细胞增殖和信号转导通路的作用。方法 体外条件下用CL-387,785处理囊肿衬里上皮细胞;四氮唑盐法(MTT)测定囊肿细胞增殖;流式细胞术检测细胞周期及凋亡;Western印迹检测EGFR磷酸化程度;细胞免疫化学方法检测核因子表达水平。结果 纳摩尔水平的CL-387,785能有效抑制细胞增殖(P<0.001),且没有明显毒性。细胞周期停滞于G_0/G_1期,未出现明显细胞凋亡。EGFR的磷酸化水平下降,核因子c-jun、c-fos表达水平显著降低(P<0.05)。结论 酪氨酸激酶抑制剂CL-387,785通过非细胞毒性作用明显抑制人多囊肾病囊肿衬里上皮细胞的增殖,其机制可能是阻断了EGFR介导的信号转导通路。Objective To investigate the effects of tyrosine kinase inhibitor CL-387,785 on the proliferation and signal transduction pathway in ADPKD cyst-lining epithelial cells. Methods Cyst-lining epithelial cells were treated by serial concentration of CL-387,785 in vitro. The proliferation was assessed by MTT. Cell cycle and apoptosis were analyzed by flow cytometry. The phosphorylation of epidermal growth factor receptor(EGFR) was detected by Western blotting and the expression of c-fos,c-jun was tested by immunocytochemistry. Results Nanomolar per liter of CL-387,785 could effectively inhibit the proliferation of the cells without obvious toxicity,and provoke G0/G1 phase arrest but could not induce apoptosis. CL-387,785 also inhibited the phosphorylation of EGFR and decreased the expression of c-fos and c-jun(P < 0.05) . Conclusions CL-387,785 can effectively block the proliferation of cyst-lining epithelial cells in a cytostatic manner. The effect may be mediated by blocking the abnormal EGFR signal transduction pathway.

关 键 词：酪氨酸激酶抑制剂 上皮细胞增殖 信号转导通路 常染色体显性遗传性多囊肾病 

分 类 号：R692.1[医药卫生—泌尿科学]