RB status as a determinant of response to UCN-01 in non-small cell lung carcinoma.
RB status as a determinant of response to UCN-01 in non-small cell lung carcinoma.
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发表时间：
1999-09
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通讯作者：
P. Mack;D. Gandara;C. Bowen;M. Edelman;T. Paglieroni;J. Schnier;E. Gelmann;P. Gumerlock
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作者：
P. Mack;D. Gandara;C. Bowen;M. Edelman;T. Paglieroni;J. Schnier;E. Gelmann;P. Gumerlock
7-羟基星孢菌素（UCN-01）是一种处于临床开发阶段的蛋白激酶抑制剂，具有很强的抗肿瘤活性。为了确定特定的遗传异常是否会调节对UCN-01的反应，这些研究使用了具有p16 CDKN 2、RB和p53差异异常的人非小细胞肺癌（NSCLC）细胞系模型。通过3-[4，5-二甲基噻唑-2-基]-2，5-二苯基溴化四唑测定来测量细胞生长，并且使用DNA含量的流式细胞术分析来研究细胞周期。蛋白质水平和磷酸化的变化通过蛋白质印迹法进行评估。在表达野生型RB的细胞系（A549和Calul）中，UCN-01处理导致剂量依赖性生长抑制、G1期细胞停滞和S期细胞减少。p16 CDKN 2-null细胞表现出与正常胎肺成纤维细胞相似的生长抑制。在p53野生型和突变型细胞系中，UCN-01诱导的生长停滞伴随着p21 CDKN 1的诱导和Rb向低磷酸化状态的转变。相反，UCN-01处理RB-无效细胞系H596导致较少的生长抑制。为了测试RB在响应UCN-01中的作用，在RB表达的两种人等基因模型中检查了治疗效果：膀胱癌细胞系5637（RB-无效）和前列腺癌细胞系DU-145（RB-突变体）。在Rb表达的5637亚系（RB 5）中，UCN-01处理导致Rb磷酸化不足，并在G1中积累，与亲本系相反。类似地，野生型Rb表达DU-145亚系（DU1.1和B5）显示与亲本细胞相比G1期阻滞增加。我们的结论是，UCN-01诱导的G1期阻滞可以发生在p53和p16 CDKN 2无效的细胞中，RB状态影响UCN-01诱导G1期阻滞的能力。这些数据表明，细胞周期调控基因在个体肿瘤的分子谱可以预测反应性，并提供洞察这种新的抗肿瘤药物的最佳治疗应用。
7-Hydroxystaurosporine (UCN-01), a protein kinase inhibitor in clinical development, demonstrates potent antineoplastic activity. To determine whether specific genetic abnormalities would modulate the response to UCN-01, a model of human non-small cell lung carcinoma (NSCLC) cell lines with differential abnormalities of p16CDKN2, RB, and p53 was used for these studies. Cell growth was measured by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and cell cycling was studied using flow cytometric analysis of DNA content. Changes in protein levels and phosphorylation were assessed by Western blotting. In cell lines expressing wild-type RB (A549 and Calul), UCN-01 treatment resulted in dose-dependent growth inhibition, arrest of cells in G1, and a reduction of cells in S phase. p16CDKN2-null cells showed similar growth inhibition to normal fetal lung fibroblasts. UCN-01-induced growth arrest was accompanied by induction of p21CDKN1 and a shift of Rb to the hypophosphorylated state in both p53 wild-type and mutant cell lines. In contrast, UCN-01 treatment of the RB-null cell line H596 resulted in less growth inhibition. To test the role of RB in response to UCN-01, effects of treatment were examined in two human isogenic models of RB expression: the bladder cancer cell line 5637 (RB-null) and the prostate cancer cell line DU-145 (RB-mutant). In the Rb-expressing 5637 subline (RB5), UCN-01 treatment resulted in Rb hypophosphorylation and an accumulation in G1 in contrast to the parent line. Similarly, the wild-type Rb-expressing DU-145 sublines (DU1.1 and B5) showed increased G1 arrest compared with the parent cells. We conclude that UCN-01-induced G1 arrest can occur in cells null for p53 and p16CDKN2, and that RB status influences the ability of UCN-01 to induce a G1 arrest. These data suggest that the molecular profile of cell cycle regulating genes in individual tumors may predict responsiveness and provide insight into optimal therapeutic application of this new antineoplastic agent.