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通过使用非特异性激酶抑制剂staurosporine(5μg·L-1),揭示正常与肿瘤细胞之间在G1/S时相过渡阶段调控的不同.方法:流式细胞光度术,点杂交,激酶活性分析,电泳.结果:18小时staurosporine(5μg·L-1)的处理,可以阻断正常细胞系2BS细胞(人胚胎肺成纤维细胞,5-20代)于G1期,降低其胞内胸腺嘧啶核苷激酶(TK)的转录和活性水平,同时也去磷酸化胞内一个107kDa的蛋白.在2BS细胞系中,所有以上现象在洗去staurosporine后都可以得到逆转.但是,所有以上结果在BGC823细胞内(人胃癌细胞)未曾出现.结论:在G1/S时相过渡阶段,正常与肿瘤细胞的调节存在差别.在正常细胞中,参与这一调节的激酶对staurosporine(5μg·L-1)比在肿瘤细胞内的激酶敏感得多.
By using the nonspecific kinase inhibitor staurosporine (5μg · L-1), the difference between normal and tumor cells in G1 / S phase transition was revealed. Methods: Flow cytometry, dot blot, kinase activity assay, electrophoresis. Results: Treatment with 18 hours staurosporine (5μg · L-1) blocked the normal cell line 2BS cells (human embryonic lung fibroblasts, 5-20 passages) in G1 phase and reduced their intracellular thymidine kinase ( TK) transcription and activity levels, but also to dephosphorylate a 107kDa intracellular protein. In the 2BS cell line, all of the above can be reversed after staurosporine washout. However, all the above results in BGC 823 cells (human gastric cancer cells) did not occur. CONCLUSIONS: There is a difference between normal and tumor cell regulation during the G1 / S phase transition. In normal cells, the kinases involved in this regulation are much more sensitive to staurosporine (5 μg · L -1) than those in tumor cells.