摘要
目的研究二甲基阿米洛利(dimethyl amiloride,DMA)对人高转移巨细胞肺癌细胞PGCL3增殖的作用及对细胞周期相关蛋白的影响。方法不同浓度的DMA作用于PGCL3细胞,用MTT法检测对细胞增殖的抑制作用;流式细胞术检测DMA对PGCL3细胞周期的影响;Westernblot检测DMA对细胞p21、cyclinA、细胞周期依赖激酶2(cell cycle depend on kinas e2,CDK2)、cyclinB1和细胞分裂周期基因25B(cell divide cycle 25B,Cdc25B)蛋白表达以及CDK2和Cdc25B蛋白活性的影响。结果DMA可抑制PGCL3细胞的增殖,并呈时间和浓度依赖性。DMA阻滞细胞于G2/M期和S期,上调p21蛋白表达,下调cyclinB1蛋白表达,降低CDK2和Cdc25B蛋白活性,但对cyclinA、CDK2和Cdc25B蛋白表达水平无影响。结论DMA可抑制人肺癌细胞增殖,并改变某些细胞周期相关蛋白的表达和活性,这可能是其抑制肿瘤的机制之一。
【Objective】To study the effect of dimethyl amiloride（DMA） on the proliferation and the expression of cell cycle-related proteins of PGCL3 cells from a human highly metastatic giant-cell lung carcinoma cell line in vitro.【Methods】PGCL3 cells were incubated with DMA at various concentration, MTT assay was used to determine the growth inhibition by DMA in PGCL3 cells; flow cytometry was applied to detect the change in cell cycle; the expression level of p21, cyclinA, CDK2 （cell cycle depend on kinase 2）, cyclinB1 and Cdc25B （cell divide cycle 25B） protein and the activity of CDK1 and Cdc25B were assessed by Western blot.【Results】DMA inhibited the proliferation of human lung carcinoma cells PGCL3 on a time-and concentration-dependent manner; DMA treatment arrested cell cycle in G2/M phase and S phase; after 24h treatment, DMA up-regulated the expression of p21 protein, down-regulated the expression of cyclinB1 protein, reduced the activity of CDK2 and Cdc25B, but did not affect the the expression of cyclinA, CDK2 and Cdc25. 【Conclusion】DMA can inhibite the proliferation of PGCL3 cells, the effect of DMA on changing the expression and activity of cell cycle-related proteins might be one of the mechanisms underlying tumor suppressive effeccts of DMA.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2010年第5期684-688,共5页
China Journal of Modern Medicine
基金
广东省医学科研基金(No:B2008086)
广东药学院博士启动基金(No:2006SMK03)
关键词
二甲基阿米洛利
肺癌
细胞增殖
细胞周期
dimethyl amiloride
lung neoplasms
cell proliferation
cell cycle
