Inhibition of epidermal growth factor receptor kinase induces protease-dependent apoptosis in human colon cancer cells
Inhibition of epidermal growth factor receptor kinase induces protease-dependent apoptosis in human colon cancer cells
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DOI：
10.1016/s0016-5085(98)70312-9
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发表时间：
1998-05-01
期刊：
影响因子：
29.4
通讯作者：
Kaufmann, SH
中科院分区：
文献类型：
--
作者：
Karnes, WE;Weller, SG;Kaufmann, SH
背景与目的：表皮生长因子受体（epidermal growth factor receptor，EGFR）作为肿瘤的治疗靶点正在研究中。结肠癌细胞系依赖于EGFR的自分泌刺激。因此，我们研究了选择性EGFR酪氨酸激酶抑制剂PD 153035对5种结肠癌细胞系增殖和存活的影响，这些细胞系的自主增殖是EGFR配体依赖性的或EGFR配体独立性的。研究方法：通过MTS生长测定、[H-3]胸苷掺入、末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记测定、荧光显微镜、免疫印迹和体外蛋白酶测定筛选抑制剂的作用。结果如下：PD 153035在配体依赖性细胞系中引起剂量依赖性细胞停滞（200 nmol/L至1 μ mol/L）和细胞凋亡（>10 μ mol/L），在配体非依赖性细胞系中引起可变细胞凋亡（>10 μ mol/L），但无细胞停滞。10 μ mol/L PD 153035诱导的细胞凋亡与p53蛋白表达的诱导无关，但伴随着切割聚（ADP-核糖）聚合酶、核纤层蛋白B-1和Bcl-2的半胱天冬酶的激活。DEVD-氟甲基酮抑制caspase 3样蛋白酶活性可显著延迟PD 153035诱导的细胞凋亡的发生。结论：EGFR酪氨酸激酶抑制剂PD 153035在EGFR配体依赖性结肠癌细胞系中诱导细胞停滞和半胱天冬酶依赖性凋亡。这些观察结果鼓励进一步研究EGFR酪氨酸激酶抑制剂治疗结直肠肿瘤。
Background & Aims: The epidermal growth factor receptor (EGFR) is under investigation as a therapeutic target for cancers. Colon cancer cell lines are variably dependent on autocrine stimulation of EGFR. We therefore examined the effects of a selective EGFR tyrosine kinase inhibitor, PD 153035, on proliferation and survival of five colon cancer cell lines whose autonomous proliferation is either EGFR ligand dependent or EGFR ligand independent. Methods: Effects of inhibitors were screened by MTS growth assays, [H-3]thymidine incorporation, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay, fluorescence microscopy, immunoblotting, and in vitro protease assays. Results: PD 153035 caused dose-dependent cytostasis (200 nmol/L to 1 mu mol/L) and apoptosis (>10 mu mol/L) in ligand-dependent cell lines and caused variable apoptosis (>10 mu mol/L) but no cytostasis in ligand-independent cell lines. Apoptosis induced by 10 mu mol/L PD 153035 was not associated with induction of p53 protein expression but was accompanied by activation of caspases that cleave poly(ADP-ribose) polymerase, lamin B-1, and Bcl-2. Inhibition of caspase 3-like protease activity by DEVD-fluoromethylketone significantly delayed the onset of PD 153035-induced apoptosis. Conclusions: The EGFR tyrosine kinase inhibitor PD 153035 induces cytostasis and caspase-dependent apoptosis in EGFR ligand-dependent colon cancer cell lines. These observations encourage further investigation of EGFR tyrosine kinase inhibitors for treatment of colorectal neoplasms.