Chen Ying, Wang Seu-Mei, Wu Jiahn-Chun, Huang Shih-Horng
Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.
J Cell Biochem. 2006 Jul 1;98(4):1021-35. doi: 10.1002/jcb.20839.
Peroxisome proliferator-activated receptor gamma (PPARgamma) agonists cause cell death in several types of cancer cells. The aim of this study was to examine the effects of two PPARgamma agonists, ciglitazone and 15-deoxy-delta(12,14)-prostaglandin J2 (15dPGJ2), on the survival of thyroid carcinoma CGTH W-2 cells. Both ciglitazone and 15dPGJ2 decreased cell viability in a time- and dose-dependent manner. Cell death was mainly due to apoptosis, with a minor contribution from necrosis. Increased levels of active caspase 3, cleaved poly (ADP-ribose) polymerase (PARP), and cytosolic cytochrome-c were noted. In addition, ciglitazone and 15dPGJ2 induced detachment of CGTH W-2 cells from the culture substratum. Both the protein levels and immunostaining signals of focal adhesion (FA) proteins, including vinculin, integrin beta1, focal adhesion kinase (FAK), and paxillin were decreased after PPARgamma agonist treatment. Meanwhile, reduced phosphorylation of FAK and paxillin was noted. Furthermore, PPARgamma agonists induced expression of protein tyrosine phosphatase-PEST (PTP-PEST), and of phosphatase and tensin homologue deleted on chromosome ten (PTEN). The upregulation of these phosphatases might contribute to the dephosphorylation of FAK and paxillin, since pre-treatment with orthovanadate prevented PPARgamma agonist-induced dephosphorylation of FAK and paxillin. Perturbation of CGTH W-2 cells with anti-integrin beta1 antibodies induced FA disruption and apoptosis in the same cells, thus the downregulation of integrin beta1 by PPARgamma agonists resulted in FA disassembly and might induce apoptosis via anoikis. Our results suggested the presence of crosstalk between apoptosis and integrin-FA signaling. Moreover, upregulation and activation of PTEN was correlated with reduced phosphorylation of Akt, and this consequence disfavored cell survival. In conclusion, PPARgamma agonists induced apoptosis of thyroid carcinoma cells via the cytochrome-c caspase 3 and PTEN-Akt pathways, and induced necrosis via the PARP pathway.
过氧化物酶体增殖物激活受体γ(PPARγ)激动剂可导致多种癌细胞死亡。本研究旨在探讨两种PPARγ激动剂,即环格列酮和15-脱氧-Δ¹²,¹⁴-前列腺素J2(15dPGJ2)对甲状腺癌CGTH W-2细胞存活的影响。环格列酮和15dPGJ2均以时间和剂量依赖性方式降低细胞活力。细胞死亡主要归因于凋亡,坏死起较小作用。观察到活性半胱天冬酶3、裂解的聚(ADP-核糖)聚合酶(PARP)和细胞溶质细胞色素c水平升高。此外,环格列酮和15dPGJ2诱导CGTH W-2细胞从培养基质上脱离。PPARγ激动剂处理后,包括纽蛋白、整合素β1、粘着斑激酶(FAK)和桩蛋白在内的粘着斑(FA)蛋白的蛋白质水平和免疫染色信号均降低。同时,观察到FAK和桩蛋白的磷酸化减少。此外,PPARγ激动剂诱导蛋白酪氨酸磷酸酶-PEST(PTP-PEST)以及10号染色体上缺失的磷酸酶和张力蛋白同源物(PTEN)的表达。这些磷酸酶的上调可能导致FAK和桩蛋白的去磷酸化,因为用原钒酸盐预处理可阻止PPARγ激动剂诱导的FAK和桩蛋白去磷酸化。用抗整合素β1抗体干扰CGTH W-2细胞可诱导同一细胞中的FA破坏和凋亡,因此PPARγ激动剂导致的整合素β1下调导致FA解体,并可能通过失巢凋亡诱导凋亡。我们的结果表明凋亡与整合素-FA信号之间存在串扰。此外,PTEN的上调和激活与Akt磷酸化减少相关,这一结果不利于细胞存活。总之,PPARγ激动剂通过细胞色素c-半胱天冬酶3和PTEN-Akt途径诱导甲状腺癌细胞凋亡,并通过PARP途径诱导坏死。