Differential effects of staurosporine analogues on cell cycle, growth and viability in A549 cells
Differential effects of staurosporine analogues on cell cycle, growth and viability in A549 cells
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DOI：
10.1038/bjc.1996.517
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发表时间：
1996-10-01
影响因子：
8.8
通讯作者：
Gescher, A
中科院分区：
文献类型：
--
作者：
Courage, C;Snowden, R;Gescher, A
Staurosporine 是一种有效但非特异性激酶抑制剂。它已作为多种对蛋白激酶 C (PKC) 具有高特异性的类似物的合成模板。此处，星形孢菌素和四种 PKC 选择性类似物（吲哚咔唑、UCN-01 和 CGP 41251，以及双吲哚基马来酰亚胺、Ro 31-8220 和 GF 109203X）作为人源 A549 人肺腺癌细胞的生长抑制剂进行了研究。比较了它们的以下方面：(1) 对细胞周期的影响、(2) 生长停滞的时间依赖性和 (3) 细胞毒性效力。将细胞暴露于星孢菌素类似物1、2和4天，或者在周期同步细胞的情况下暴露6、12和24小时，其浓度在暴露4天后抑制生长80%。 Staurosporine 和 UCN-01 使细胞延迟 G(0/1)，而 CCP 41251 似乎抑制细胞生长，但没有细胞周期特异性。 Ro 31-8220 减缓了周期中同步细胞的进展；在较长一段时间内，它会引起 G(2)/M 的弱阻滞。 GF 109203X 在同步细胞中诱导有效的 G(2)/M 停滞。这在异步细胞中并不那么明显，到了第 4 天，异步细胞在 G(0/1) 中减慢了速度。这些抑制剂诱导的生长停滞在孵育 4 天而不是 2 天后更有效。孵育 1 天，然后在无药物培养基中维持 3 天足以发挥一定的细胞抑制作用。细胞毒性浓度和细胞抑制浓度之间的差异（前者通过乳酸脱氢酶的细胞释放来测量）对于星形孢菌素来说是15000倍，对于UCN-01来说是300倍，对于CGP 41251来说是400倍，对于Ro 31-8220来说是25倍，对于GF 109203X来说是4倍。结果表明，PKC 选择性星形孢菌素类似物在干扰细胞周期的机制方面有所不同。长期暴露以有效抑制生长的必要性以及细胞抑制浓度和急性细胞毒性吲哚并咔唑浓度之间的相当大的裕度可能会影响这些激酶抑制剂临床试验的计划和解释。
Staurosporine is a potent but non-specific kinase inhibitor. It has served as synthetic template for a variety of analogues with high specificity for protein kinase C (PKC). Here staurosporine and four PKC-selective analogues, the indolocarbazoles, UCN-01 and CGP 41251, and the bisindolylmaleimides, Ro 31-8220 and GF 109203X, were investigated as growth inhibitors of human-derived A549 human lung adenocarcinoma cells. They were compared with respect to (1) effect on the cell cycle, (2) time dependency of growth arrest and (3) cytotoxic potency. Cells were exposed for 1, 2 and 4 days, or for 6, 12 and 24 h in the case of cycle-synchronised cells, to staurosporine analogues at concentrations at which they inhibited growth by 80% after 4 day exposure. Staurosporine and UCN-01 retarded cells in G(0/1), and CCP 41251 appeared to inhibit cell growth without cell cycle specificity. Ro 31-8220 slowed progression of synchronised cells through the cycle; over a longer time period it induced a weak block in G(2)/M. GF 109203X induced potent G(2)/M arrest in synchronised cells. This was not so apparent in asynchronous cells, which by day 4 were slowed in G(0/1) instead. Growth arrest induced by these inhibitors was more potent after incubation for 4 rather than 2 days. Incubation for 1 day followed by maintenance in drug-free medium for 3 days was sufficient to exert some cytostasis. The differences between cytotoxic and cytostatic concentrations, the former measured by release from cells of lactate dehydrogenase, were 15 000-fold for staurosporine, 300-fold for UCN-01, similar to 400-fold for CGP 41251, 25-fold for Ro 31-8220 and similar to 4-fold for GF 109203X. The results show that PKC-selective staurosporine analogues differ with respect to the mechanisms by which they interfere with the cell cycle. The necessity of long-term exposure for effective growth inhibition and the considerable margin between cytostatic and acute cytotoxic indolocarbazole concentrations are findings which might influence the planning and interpretation of clinical trials of these kinase inhibitors.