Kawakami Masanori, Mustachio Lisa Maria, Rodriguez-Canales Jaime, Mino Barbara, Roszik Jason, Tong Pan, Wang Jing, Lee J Jack, Myung Ja Hye, Heymach John V, Johnson Faye M, Hong Seungpyo, Zheng Lin, Hu Shanhu, Villalobos Pamela Andrea, Behrens Carmen, Wistuba Ignacio, Freemantle Sarah, Liu Xi, Dmitrovsky Ethan
Departments of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
J Natl Cancer Inst. 2017 Jun 1;109(6). doi: 10.1093/jnci/djw297.
The first generation CDK2/7/9 inhibitor seliciclib (CYC202) causes multipolar anaphase and apoptosis in lung cancer cells with supernumerary centrosomes (known as anaphase catastrophe). We investigated a new and potent CDK2/9 inhibitor, CCT68127 (Cyclacel).
CCT68127 was studied in lung cancer cells (three murine and five human) and control murine pulmonary epithelial and human immortalized bronchial epithelial cells. Robotic CCT68127 cell-based proliferation screens were used. Cells undergoing multipolar anaphase and inhibited centrosome clustering were scored. Reverse phase protein arrays (RPPAs) assessed CCT68127 effects on signaling pathways. The function of PEA15, a growth regulator highlighted by RPPAs, was analyzed. Syngeneic murine lung cancer xenografts (n = 4/group) determined CCT68127 effects on tumorigenicity and circulating tumor cell levels. All statistical tests were two-sided.
CCT68127 inhibited growth up to 88.5% (SD = 6.4%, P < .003) at 1 μM, induced apoptosis up to 42.6% (SD = 5.5%, P < .001) at 2 μM, and caused G1 or G2/M arrest in lung cancer cells with minimal effects on control cells (growth inhibition at 1 μM: 10.6%, SD = 3.6%, P = .32; apoptosis at 2 μM: 8.2%, SD = 1.0%, P = .22). A robotic screen found that lung cancer cells with KRAS mutation were particularly sensitive to CCT68127 ( P = .02 for IC 50 ). CCT68127 inhibited supernumerary centrosome clustering and caused anaphase catastrophe by 14.1% (SD = 3.6%, P < .009 at 1 μM). CCT68127 reduced PEA15 phosphorylation by 70% (SD = 3.0%, P = .003). The gain of PEA15 expression antagonized and its loss enhanced CCT68127-mediated growth inhibition. CCT68127 reduced lung cancer growth in vivo ( P < .001) and circulating tumor cells ( P = .004). Findings were confirmed with another CDK2/9 inhibitor, CYC065.
Next-generation CDK2/9 inhibition elicits marked antineoplastic effects in lung cancer via anaphase catastrophe and reduced PEA15 phosphorylation.
第一代细胞周期蛋白依赖性激酶2/7/9（CDK2/7/9）抑制剂塞利西利布（CYC202）可在具有多余中心体的肺癌细胞中引发多极后期和细胞凋亡（即后期灾难）。我们研究了一种新型强效CDK2/9抑制剂CCT68127（Cyclacel）。
在肺癌细胞（三种小鼠肺癌细胞和五种人肺癌细胞）以及对照小鼠肺上皮细胞和人永生化支气管上皮细胞中研究CCT68127。采用基于机器人的CCT68127细胞增殖筛选。对经历多极后期和中心体聚集受抑制的细胞进行评分。反向蛋白质阵列（RPPA）评估CCT68127对信号通路的影响。分析了RPPA突出显示的生长调节因子PEA15的功能。同基因小鼠肺癌异种移植瘤（每组n = 4）确定CCT68127对肿瘤发生性和循环肿瘤细胞水平的影响。所有统计检验均为双侧检验。
CCT68127在1 μM时可抑制生长达88.5%（标准差= 6.4%，P <.003），在2 μM时可诱导细胞凋亡达42.6%（标准差= 5.5%，P <.001），并在肺癌细胞中引起G1期或G2/M期阻滞，而对对照细胞影响极小（1 μM时生长抑制率：10.6%，标准差= 3.6%，P = 0.32；2 μM时细胞凋亡率：8.2%，标准差= 1.0%，P = 0.22）。一项机器人筛选发现，具有KRAS突变的肺癌细胞对CCT68127特别敏感（IC50的P值为0.02）。CCT68127抑制多余中心体聚集，并在1 μM时导致14.1%的后期灾难（标准差= 3.6%，P <.009）。CCT68127使PEA15磷酸化降低70%（标准差= 3.0%，P = 0.003）。PEA15表达增加可拮抗，而其缺失则增强CCT68127介导的生长抑制。CCT68127在体内可降低肺癌生长（P <.001）和循环肿瘤细胞水平（P = 0.004）。另一项研究使用另一种CDK2/9抑制剂CYC065证实了这些发现。
新一代CDK2/9抑制通过后期灾难和降低PEA15磷酸化在肺癌中引发显著的抗肿瘤作用。
