Yang Lin, Shu Taipengfei, Liang Yingjian, Gu Wenguang, Wang Chunlei, Song Xuanhe, Fan Changdong, Wang Wenbo
Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.
Department of Endocrinology, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.
Int J Oncol. 2015 Apr;46(4):1651-8. doi: 10.3892/ijo.2015.2872. Epub 2015 Feb 4.
Angiopoietin-like protein 2 (ANGPTL2) plays an important role in inflammatory carcinogenesis and tumor metastasis. The compound GDC-0152 is a peptidomimetic small molecule antagonist of inhibitor of apoptosis (IAP) proteins with antitumor activity. However, the interaction between ANGPTL2 and GDC-0152 has not been studied. It has been proven that ANGPTL2 promotes metastasis of osteosarcoma. Therefore, in the present study, the effect of GDC-0152 on the malignant progression of osteosarcoma promoted by ANGPTL2 was investigated. Human osteosarcoma cell line SaOS2 cells were pre-treated or non-treated with GDC-0152 and then exposed to recombinant human ANGPTL2. The viability of SaOS2 cells was determined by MTT assay, the migration of SaOS2 cells was analyzed by chamber migration assay kit, and the SaOS2 cell apoptosis was determined by fluorescence-activated cell sorting (FACS) and nuclear staining. Treatment with ANGPTL2 increased SaOS2 cell growth and migration and decreased cell apoptosis. The increased cell growth and decreased cell apoptosis were significantly attenuated in SaOS2 cells receiving GDC-0152. However, the ANGPTL2-increased SaOS2 cell migration was not inhibited by GDC-0152 treatment. Furthermore, western blot analysis showed that the activation of phosphatidyl inositol 3-kinase (PI3K) (p85), PI3K (p110), protein kinase B (Akt) (Ser473), Akt (Thr308) and p38 mitogen-activated protein kinase (p38MAPK) were upregulated by ANGPTL2. Quantitative real-time polymerase chain reaction (qTR-PCR) and gelatin zymography showed that the mRNA expression and activity of matrix metalloproteinase-9 (MMP-9) and matrix metalloproteinase-2 (MMP-2) were also increased by ANGPTL2. The upregulated activation of PI3K and Akt were significantly suppressed by the treatment of GDC-0152. In contrast, GDC-0152 did not suppress ANGPTL2-induced p38MAPK phosphorylation, MMP-9/MMP-2 mRNA expression or MMP-9/MMP-2 activity. Taken together, these data indicate that GDC-0152 attenuates the malignant progression of osteosarcoma promoted by ANGPTL2 via PI3K/AKT but not p38MAPK signaling pathway. The present study indicated a novel therapeutic strategy to inhibit tumor growth by indirectly preventing ANGPTL2 signaling.
血管生成素样蛋白2（ANGPTL2）在炎症致癌和肿瘤转移中起重要作用。化合物GDC - 0152是一种具有抗肿瘤活性的凋亡抑制蛋白（IAP）的拟肽小分子拮抗剂。然而，ANGPTL2与GDC - 0152之间的相互作用尚未得到研究。已证实ANGPTL2促进骨肉瘤转移。因此，在本研究中，研究了GDC - 0152对ANGPTL2促进的骨肉瘤恶性进展的影响。人骨肉瘤细胞系SaOS2细胞用GDC - 0152预处理或未处理，然后暴露于重组人ANGPTL2。通过MTT法测定SaOS2细胞的活力，用Transwell迁移实验试剂盒分析SaOS2细胞的迁移情况，通过荧光激活细胞分选（FACS）和细胞核染色测定SaOS2细胞凋亡。ANGPTL2处理增加了SaOS2细胞的生长和迁移，并降低了细胞凋亡。在接受GDC - 0152的SaOS2细胞中，细胞生长增加和细胞凋亡减少的情况明显减弱。然而，GDC - 0152处理并未抑制ANGPTL2增加的SaOS2细胞迁移。此外，蛋白质印迹分析表明，磷脂酰肌醇3激酶（PI3K）（p85）、PI3K（p110）、蛋白激酶B（Akt）（Ser473）、Akt（Thr308）和p38丝裂原活化蛋白激酶（p38MAPK）的激活被ANGPTL2上调。定量实时聚合酶链反应（qRT - PCR）和明胶酶谱分析表明，基质金属蛋白酶9（MMP - 9）和基质金属蛋白酶2（MMP - 2）的mRNA表达和活性也被ANGPTL2上调。GDC - 0152处理显著抑制了PI3K和Akt的上调激活。相反，GDC - 0152并未抑制ANGPTL2诱导的p38MAPK磷酸化、MMP - 9/MMP - 2 mRNA表达或MMP - 9/MMP - 2活性。综上所述，这些数据表明GDC - 0152通过PI3K/AKT而非p38MAPK信号通路减弱了ANGPTL2促进的骨肉瘤恶性进展。本研究表明了一种通过间接阻断ANGPTL2信号来抑制肿瘤生长的新治疗策略。
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