Hishiki Tomoro, Saito Takeshi, Sato Yoshiharu, Mitsunaga Tetsuya, Terui Elena, Matsuura Gen, Saito Eriko, Shibata Ryohei, Mise Naoko, Yokoyama Yukiko, Yoshida Hideo
Department of Pediatric Surgery, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8677, Japan.
Pediatr Surg Int. 2011 Feb;27(2):225-30. doi: 10.1007/s00383-010-2775-2.
Neuroblastoma (NB) is one of the most common extracranial solid tumors in children and is known for its clinical and biological heterogeneity. The aim of this study is to reveal the functional role of src family kinases in the biological behavior of NB by inhibiting their kinase activities with a specific inhibitor, PP2 (4-amino-5-(4-chloro-phenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine).
NB cell lines (SH-SY5Y, IMR32, RT-BM-1, CHP134, NLF, and LA-N-5) were treated with 0.1-10 µM of PP2. Morphological changes, cell growth, and cell death were assessed, as well as all-trans retinoic acid (ATRA)-induced neuronal differentiation and epidermal growth factor (EGF)-induced proliferation.
At 24 h after PP2 treatment, NB cell lines showed drastic cell aggregation. PP2 also inhibited cell growth of NB in a dose-dependent manner. Apoptosis was detected in these cells. ATRA-induced neuronal differentiation of RT-BM-1 was not affected by PP2. PP2 reduced the proliferative effect of EGF. EGF-induced rapid activation of Akt, which was not blocked by PP2 treatment, suggesting that the cellular events triggered by PP2 were independent to PI3 kinase/Akt signaling pathway.
Our data suggests that src family kinases promote cell survival/proliferation and reduces cell aggregation of NBs. Src family kinase inhibitors may be good candidates for a novel molecular target therapy.
神经母细胞瘤（NB）是儿童最常见的颅外实体瘤之一，以其临床和生物学异质性而闻名。本研究的目的是通过用特异性抑制剂PP2（4-氨基-5-（4-氯苯基）-7-（叔丁基）吡唑并[3,4-d]嘧啶）抑制src家族激酶的激酶活性，揭示其在NB生物学行为中的功能作用。
用0.1-10μM的PP2处理NB细胞系（SH-SY5Y、IMR32、RT-BM-1、CHP134、NLF和LA-N-5）。评估形态学变化、细胞生长和细胞死亡，以及全反式维甲酸（ATRA）诱导的神经元分化和表皮生长因子（EGF）诱导的增殖。
PP2处理24小时后，NB细胞系出现剧烈的细胞聚集。PP2还以剂量依赖性方式抑制NB的细胞生长。在这些细胞中检测到凋亡。PP2不影响ATRA诱导的RT-BM-1神经元分化。PP2降低了EGF的增殖作用。EGF诱导的Akt快速激活不受PP2处理的阻断，这表明PP2触发的细胞事件独立于PI3激酶/Akt信号通路。
我们的数据表明，src家族激酶促进NB的细胞存活/增殖并减少细胞聚集。Src家族激酶抑制剂可能是新型分子靶向治疗的良好候选药物。
