Effect of troglitazone on tumor growth and pulmonary metastasis development of the mouse osteosarcoma cell line LM8
Effect of troglitazone on tumor growth and pulmonary metastasis development of the mouse osteosarcoma cell line LM8
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DOI：
10.1186/1471-2407-10-51
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发表时间：
2010-02-22
期刊：
影响因子：
3.8
通讯作者：
Masuno, Hiroshi
中科院分区：
文献类型：
--
作者：
Aizawa, Junichi;Sakayama, Kenshi;Masuno, Hiroshi
背景：骨肉瘤在确诊前常发生肺部微转移，导致致命后果。因此，预防肺转移是改善骨肉瘤患者预后的关键。本研究旨在探讨曲格列酮(TGZ)是否可作为治疗骨肉瘤生长和转移的药物。用DNA测定法和5-溴-2‘-脱氧尿嘧啶核苷掺入法检测TGZ对细胞增殖的影响。在侵袭室中使用Matrigel包被的细胞培养插入物或未包被的细胞培养插入物进行细胞侵袭和运动能力的测定。通过Western印迹分析Akt和p-Akt来评价TGZ对Akt信号转导的影响。采用裸鼠背部移植LM8细胞模型，观察口服TGZ(TGZ组)和乙醇(对照组)对原发肿瘤生长和肺转移的影响。免疫组织化学和酶谱法检测肿瘤组织中基质金属蛋白酶-2(MMP2)的表达和活性。CD34免疫组织化学方法检测肿瘤内微血管密度(MVD)。结果：TGZ呈剂量依赖性抑制细胞增殖。TGZ处理的细胞比未处理的细胞侵袭性更小，运动能力更差。TGZ处理的细胞分泌的基质金属蛋白酶-2活性低于未处理的细胞。TGZ可降低p-Akt水平。TGZ组的原发肿瘤质量明显小于对照组。与对照组相比，TGZ组肺内转移瘤较少。TGZ组肿瘤内基质金属蛋白酶-2的表达和活性均低于对照组。结论：TGZ可抑制Akt信号转导途径，减少MMP2的分泌，从而降低LM8细胞的侵袭力和运动能力。TGZ治疗可降低荷瘤小鼠肿瘤内基质金属蛋白酶-2的表达和活性，抑制原发肿瘤的生长和肺转移的发展。TGZ可能为骨肉瘤的化疗提供一种新的途径。
Background: Osteosarcoma often develops micrometastases in the lung prior to diagnosis, causing a fatal outcome. Therefore, the prevention of pulmonary metastases is critical for the improvement of the prognosis of patients with osteosarcoma. The purpose of this study was to investigate whether troglitazone (TGZ) is considered as possible therapeutics in the treatment of growth and metastasis of osteosarcoma.Methods: LM8 cells were treated for 3 days with various concentrations of TGZ. The effect of TGZ on cell proliferation was determined by DNA measurement in the cultures and 5-bromo-2'-deoxyuridine incorporation study. The assay of cell invasion and motility was performed using either the Matrigel-coated cell culture inserts or the uncoated cell culture inserts in the invasion chambers. The effect of TGZ on Akt signaling was assessed by Western blot analysis of Akt and p-Akt. The effects of oral administration of either TGZ (TGZ group) or ethanol (control group) on the growth of primary tumor and the development of pulmonary metastasis were examined in nude mice implanted with LM8 cells on their backs. The expression and activity of matrix metalloproteinase 2 (MMP-2) within the tumor were determined by immunohistochemistry and zymography. The microvessel density (MVD) within the tumor was determined by immunohistochemistry for CD34.Results: TGZ dose-dependently inhibits cell proliferation. TGZ-treated cells were less invasive and less motile than untreated cells. The activity of MMP-2 secreted by TGZ-treated cells was lower than that secreted by untreated cells. TGZ decreased the level of p-Akt. The primary tumor mass was smaller in the TGZ group than in the control group. The TGZ group had less metastatic tumors in the lung compared with the control group. The expression and activity of MMP-2 within the tumor of the TGZ group were lower than those of the control group. The MVD within the tumor of the TGZ group was lower than that of the control group.Conclusions: Inhibition of Akt signaling by TGZ may decrease the secretion of MMP-2, resulting in the decrease of invasiveness and motility in LM8 cells. Treatment of tumor-bearing mice with TGZ decreases the expression and activity of MMP-2 within the tumor, and inhibits primary tumor growth and pulmonary metastasis development. TGZ may offer a new approach in chemotherapy for osteosarcoma.