Flavopiridol induces cell cycle arrest and p53-independent apoptosis in non-small cell lung cancer cell lines.
Flavopiridol induces cell cycle arrest and p53-independent apoptosis in non-small cell lung cancer cell lines.
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发表时间：
1999-10
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通讯作者：
G. Shapiro;Darlene A. Koestner;C. Matranga;B. Rollins
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作者：
G. Shapiro;Darlene A. Koestner;C. Matranga;B. Rollins
Flavopiridol是一种人工合成的黄酮类化合物，在体外和体内都能抑制肿瘤生长，是一种有效的细胞周期蛋白依赖性激酶（cdk）抑制剂，目前正在进行临床试验。在本研究中，检查了100-500 nM黄酮吡啶醇对一组非小细胞肺癌细胞系的影响。所有细胞都表达野生型视网膜母细胞瘤易感蛋白，缺乏p16 INK 4A，已知只有A549细胞表达野生型p53。在72小时的治疗过程中，flavopiridol被证明是细胞毒性的所有七个细胞系，如台盼蓝排斥，无论细胞是否积极循环。在大多数细胞周期中，细胞毒性之前或伴随着细胞周期停滞。细胞死亡导致出现亚G1 DNA含量的细胞，提示细胞凋亡，这通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记试验和通过证明半胱天冬酶靶点（包括聚（ADP-核糖）聚合酶、p21 Waf 1和p27 Kip 1）的裂解得到证实。在剂量等于或低于500 nM时，最大细胞毒性需要暴露72小时。虽然flavopiridol导致A549细胞中p53的积累，但flavopiridol介导的细胞凋亡是p53独立的，因为它在A549细胞中发生的程度相同，其中p53被HPV 16 E6表达靶向降解。这些数据表明，flavopiridol在体外对非小细胞肺癌具有活性，值得在治疗这种疾病方面继续进行临床开发。
Flavopiridol, a synthetic flavone that inhibits tumor growth in vitro and in vivo, is a potent cyclin-dependent kinase (cdk) inhibitor presently in clinical trials. In the present study, the effect of 100-500 nM flavopiridol on a panel of non-small cell lung cancer cell lines was examined. All express a wild-type retinoblastoma susceptibility protein and lack p16INK4A, and only A549 cells are known to express wild-type p53. During 72 h of treatment, flavopiridol was shown to be cytotoxic to all seven cell lines, as measured by trypan blue exclusion, regardless of whether cells were actively cycling. In most cycling cells, cytotoxicity was preceded or accompanied by cell cycle arrest. Cell death resulted in the appearance of cells with a sub-G1 DNA content, suggestive of apoptosis, which was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay and by demonstration of cleavage of caspase targets including poly(ADP-ribose) polymerase, p21Waf1, and p27Kip1. At doses at or below 500 nM, maximal cytotoxicity required 72 h of exposure. Although flavopiridol resulted in the accumulation of p53 in A549 cells, flavopiridol-mediated apoptosis was p53 independent because it occurred to the same degree in A549 cells in which p53 was targeted for degradation by HPV16E6 expression. The data indicate that flavopiridol has activity against non-small cell lung cancers in vitro and is worthy of continued clinical development in the treatment of this disease.