Placha Wojciech, Gil Dorota, Dembińska-Kieć Aldona, Laidler Piotr
Institute of Medical Biochemistry, St Kopernika 7, and Department of Clinical Biochemistry, St Kopernika 15, Jagiellonian University Medical College, 31-034 Kraków, Poland.
Melanoma Res. 2003 Oct;13(5):447-56. doi: 10.1097/00008390-200310000-00003.
The peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear receptor family of ligand-activated transcription factors. PPARgamma ligands have been shown to inhibit the growth of cells from different cancer lineages. This study was designed to evaluate the effects of PPARgamma receptor activation in human melanoma cell lines. The effects of its expression and activation on cell proliferation and apoptosis in human melanoma cell lines (early stage cancer [WM35] and metastatic tumour [A375]) were investigated. Reverse transcription-polymerase chain reaction and Western blot analysis showed that both human melanoma cell lines expressed PPARgamma mRNA and protein. Treatment of cells transfected with the luciferase gene ligated to PPAR response element constructed promoter showed that ciglitazone and prostaglandin J2 (PGJ2), selective ligands for PPARgamma, increased the luciferase activity, proving the induction of the PPARgamma reporter gene. Ciglitazone and PGJ2 inhibited melanoma cell proliferation in a dose-dependent manner. Analysis of the cellular morphology and apoptosis assayed by fluorescence microscopy after incubation of A375 cells with 10 micro M ciglitazone for 24 h indicated that this ligand not only inhibited cell proliferation but also induced apoptosis.
过氧化物酶体增殖物激活受体γ（PPARγ）是配体激活转录因子核受体家族的成员。PPARγ配体已被证明可抑制来自不同癌症谱系的细胞生长。本研究旨在评估PPARγ受体激活对人黑色素瘤细胞系的影响。研究了其表达和激活对人黑色素瘤细胞系（早期癌症[WM35]和转移性肿瘤[A375]）细胞增殖和凋亡的影响。逆转录-聚合酶链反应和蛋白质印迹分析表明，两种人黑色素瘤细胞系均表达PPARγ mRNA和蛋白质。用与PPAR反应元件构建的启动子连接的荧光素酶基因转染细胞后进行处理，结果显示，PPARγ的选择性配体罗格列酮和前列腺素J2（PGJ2）增加了荧光素酶活性，证明了PPARγ报告基因的诱导。罗格列酮和PGJ2以剂量依赖性方式抑制黑色素瘤细胞增殖。用10μM罗格列酮孵育A375细胞24小时后，通过荧光显微镜分析细胞形态和凋亡，结果表明该配体不仅抑制细胞增殖，还诱导凋亡。
