In vitro and in vivo cytotoxicity of troglitazone in pancreatic cancer.
In vitro and in vivo cytotoxicity of troglitazone in pancreatic cancer.
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DOI：
10.1186/s13046-017-0557-6
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发表时间：
2017-07-03
期刊：
影响因子：
--
通讯作者：
Okamura N
中科院分区：
文献类型：
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作者：
Fujita M;Hasegawa A;Yamamori M;Okamura N
曲格列酮（TGZ）是一种过氧化物酶体增殖激活受体γ (PPARγ)激动剂，已被研究为一种潜在的化学预防和化学治疗药物。然而，TGZ在胰腺癌中的抗肿瘤作用及其机制尚未得到广泛的研究。本研究旨在探讨TGZ对胰腺癌细胞系的体外和体内作用，以及TGZ对PPARγ依赖性、Akt和丝裂原活化蛋白激酶（MAPK）通路的作用机制。我们还评估了TGZ对细胞侵袭和迁移的影响。使用MIA Paca2和PANC-1人胰腺癌细胞系。荧光试剂检测细胞活力和caspase-3活性，并用Hoechst 33342染色观察细胞染色质凝集。western blot检测蛋白表达水平。入侵和迁移试验使用24孔室进行。用裸鼠接种MIA Paca2细胞，研究TGZ的体内抗肿瘤作用。小鼠每天口服TGZ (200 mg/kg)，连续5周，测量肿瘤体积。TGZ对两种细胞系均表现出剂量依赖性的细胞毒性，且不受PPARγ抑制剂的减弱。此外，TGZ在MIA Paca2细胞中诱导染色质凝聚，升高caspase-3活性，增加Bax/Bcl-2的相对表达。TGZ还增加了两种细胞系中Akt和MAPK （ERK/p38/JNK）的磷酸化，JNK抑制剂显著提高了MIA Paca2细胞的活力。TGZ适度抑制细胞迁移。TGZ给药可抑制MIA Paca2异种移植模型的肿瘤生长，但给药组小鼠体重与给药组无明显差异。我们首次证明了TGZ对胰腺癌的体内抗肿瘤作用，而没有明显的不良反应。TGZ诱导线粒体介导的MIA Paca2细胞凋亡，其细胞毒性作用不依赖ppar γ，并通过JNK途径发生。我们的研究结果表明，TGZ是一种潜在的治疗胰腺癌的方法，值得进一步研究其详细的机制和临床疗效。
Troglitazone (TGZ) is a peroxisome proliferator-activated receptor gamma (PPARγ) agonist that has been investigated as a potential chemopreventive and chemotherapeutic agent. However, the antitumor efficacy and mechanisms of TGZ in pancreatic cancer have not been extensively investigated. This study was performed to investigate the in vitro and in vivo effects of TGZ against pancreatic cancer cell lines, as well as its action mechanisms in terms of PPARγ dependency and the Akt and mitogen-activated protein kinase (MAPK) pathways. We also evaluated the effects of TGZ on cell invasion and migration. MIA Paca2 and PANC-1 human pancreatic cancer cell lines were used. Cell viability and caspase-3 activity were detected using fluorescent reagents, and chromatin condensation was observed after staining the cells with Hoechst 33342. Protein expression levels were detected by western blot analysis. Invasion and migration assays were performed using 24-well chambers. The in vivo antitumor effects of TGZ were investigated in nude mice inoculated with MIA Paca2 cells. Mice were orally administered TGZ (200 mg/kg) every day for 5 weeks, and tumor volumes were measured bi-dimensionally. TGZ showed dose-dependent cytotoxicity against both cell lines, which was not attenuated by a PPARγ inhibitor. Further, TGZ induced chromatin condensation, elevated caspase-3 activity, and increased Bax/Bcl-2 relative expression in MIA Paca2 cells. TGZ also increased phosphorylation of Akt and MAPK (ERK/p38/JNK) in both cell lines, and a JNK inhibitor significantly increased the viability of MIA Paca2 cells. TGZ moderately inhibited cell migration. Tumor growth in the MIA Paca2 xenograft model was inhibited by TGZ administration, while mouse body weights in the treated group were not different from those of the vehicle administration group. We demonstrated for the first time the in vivo antitumor effects of TGZ in pancreatic cancer without marked adverse effects. TGZ induced mitochondria-mediated apoptosis in MIA Paca2 cells, and its cytotoxic effects were PPARγ-independent and occurred via the JNK pathway. Our results indicate that TGZ is a potential approach for the treatment of pancreatic cancer and warrants further studies regarding its detailed mechanisms and clinical efficacy.