摘要
本研究观察白血病和卵巢癌亲代及耐药细胞系中端粒酶活性及细胞外调节蛋白激酶 (extracelluarregula tedproteinkinasesERK)磷酸化蛋白表达水平的变化 ,探讨端粒酶与ERK在白血病及卵巢癌耐药中的作用。采用MTT法评价白血病和卵巢癌亲代和耐药细胞系对HRT或DDP的敏感性 ,用流式细胞术分析这两种细胞系间细胞周期分布的差别 ,用端粒酶重复扩增技术 (TRAP)、生物发光分析法定量和定性检测端粒酶活性及用Westernblot检测法检测磷酸化ERK1 和ERK2 蛋白的表达水平。结果表明 :白血病和卵巢癌耐药细胞系处于G0 G1 期的细胞比例增加 ,端粒酶活性和磷酸化ERK1 2 蛋白表达水平耐药细胞系较亲代细胞系高。结论 :耐药细胞系G0 G1期细胞比例增加可能是细胞产生耐药的一种标志。白血病和卵巢癌细胞系耐药的产生可能与端粒酶活性和磷酸化ERK1 2 蛋白表达水平的增高有关。
In order to study the role of telomerase and extracelluar regulated protein kinase (ERK) in drug-resistance of leukemia and ovarian cancer cells,telomeric repeat amplification protocol (TRAP) assay and bioluminescence analysis were used for qualitative analysis or quantitive detection of telomerase activity respectively,and Western blot was used to detect the expression level of phosphorylatedly activated ERK 1 and ERK 2 protein in the parental and drug resistant cells of leukemia and ovarian cancer. In addition,chemotherapy sensitivity to HRT or DDP was evaluated by MTT assay. The difference of cell cycle distribution between parental cell and drug-resistant cell was analyzed by flow cytometry. The results showed that the drug resistant cells were of higher percentage in G 0/G 1 phase compared with the parental cell lines. Telomerase activity and phosphorylatedly activated ERK 1 and ERK 2 protein expression level were higher in drug-resistant cells than in parental cell. It is suggested that the increasing number of the drug resistant cells in G 0/G 1 phase may be considered as a sign of drug resistance. The up-regulation of telomerase activity and phosphorylatedly activated ERK 1 and ERK 2 protein expression level may play an important role in drug resistance of leukemia and ovarian cancer cell lines.
出处
《中国实验血液学杂志》
CAS
CSCD
2004年第3期304-308,共5页
Journal of Experimental Hematology
关键词
白血病
卵巢癌
耐药
端粒酶
细胞外调节蛋白激酶
leukemia
ovarian cancer
drug resistance
telomerase
extracelluar regulated protein kinases
