Bhonde Mandar R, Hanski Marie-Luise, Magrini Roberta, Moorthy Dhatchana, Müller Antje, Sausville Edward A, Kohno Kimitoshi, Wiegand Peter, Daniel Peter T, Zeitz Martin, Hanski Christoph
Department of Gastroenterology, Charité-Universitaetsmedizin Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12200 Berlin, Germany.
Oncogene. 2005 Jan 6;24(1):148-56. doi: 10.1038/sj.onc.1207842.
The broad-range cyclin-dependent kinase inhibitor 7-hydroxystaurosporine (UCN-01) is known to induce both a G1 cell cycle arrest and apoptosis. The mechanism of UCN-01-induced apoptosis is largely unknown. We analysed the mechanism of cytotoxicity of UCN-01 in four established colon carcinoma cell lines. The cell lines SW48 and LS513 responded to UCN-01 treatment by undergoing apoptosis in a concentration-dependent manner while the cell lines HT-29 and WiDr were completely resistant. Apoptosis in LS513 and SW48 cell lines was concomitant with the suppression of Bcl-x(L) on mRNA and protein level. In contrast, in the apoptosis-resistant cell lines, Bcl-x(L) expression was not affected by UCN-01. Stable overexpression of the Bcl-x(L) protein abrogated UCN-01-triggered apoptosis, but only partially restored growth, indicating that both cell cycle arrest and apoptosis exert the anticancer effect in a coordinated manner. The inhibition of Akt phosphorylation did not correlate with the apoptotic phenotype. UCN-01 inhibited the activating STAT3 phosphorylations on Ser727 and, notably, on Tyr705, but STAT3 did not contribute to Bcl-x(L) expression in colon carcinoma cells. Moreover, we show for the first time that UCN-01 induces apoptosis by suppression of Bcl-x(L) expression. The inhibition of this pathway is a new aspect of cytotoxic and modulatory potential of UCN-01.
广谱细胞周期蛋白依赖性激酶抑制剂7-羟基星孢菌素(UCN-01)已知可诱导G1期细胞周期停滞和凋亡。UCN-01诱导凋亡的机制 largely未知。我们分析了UCN-01在四种已建立的结肠癌细胞系中的细胞毒性机制。SW48和LS513细胞系对UCN-01处理的反应是呈浓度依赖性地发生凋亡,而HT-29和WiDr细胞系则完全耐药。LS513和SW48细胞系中的凋亡与Bcl-x(L)在mRNA和蛋白水平的抑制相伴。相反,在抗凋亡细胞系中,Bcl-x(L)表达不受UCN-01影响。Bcl-x(L)蛋白的稳定过表达消除了UCN-01触发的凋亡,但仅部分恢复生长,表明细胞周期停滞和凋亡均以协调的方式发挥抗癌作用。Akt磷酸化的抑制与凋亡表型无关。UCN-01抑制了Ser727以及尤其是Tyr705上的STAT3磷酸化激活,但STAT3在结肠癌细胞中对Bcl-x(L)表达没有贡献。此外,我们首次表明UCN-01通过抑制Bcl-x(L)表达诱导凋亡。该途径的抑制是UCN-01细胞毒性和调节潜力的一个新方面。