Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Perugia, Italy.
Prostate. 2013 Jan;73(2):121-32. doi: 10.1002/pros.22547. Epub 2012 Jun 1.
Glyoxalase I (GLOI) detoxifies reactive dicarbonyls, as methylglyoxal (MG) that, directly or through the formation of MG-derived adducts, is a growth inhibitor and apoptosis inducer. GLOI has been considered a general marker of cell proliferation, but a direct link between the two has yet to be demonstrated. The aim of the present work was to clarify whether GLOI was involved in the proliferation control of LNCaP and PC3 human prostate cancer cells or might play a different role in the growth regulation of these cells.
RNA interference was used to study the role of GLOI in cell proliferation or apoptosis. Cell proliferation was evaluated by [3H]thymidine incorporation assay and flow cytometry, that was also used to analyze apoptosis. Real-time TaqMan polymerase chain reaction and spectrophotometric analyses were used to study transcript levels or specific activity, respectively. Proteins levels were analyzed by Western blot. MG was measured by high-performance liquid chromatography.
We found that GLOI is not implicated in the proliferation control of LNCaP and PC3 cells but plays a role in the apoptosis of invasive prostate cancer PC3 cells, through a mechanism involving a specific MG-adduct and NF-kB signaling pathway.
Our data represent the first systematic demonstration that GLOI cannot be considered a general marker of cell proliferation and that acts as a pro-survival factor in invasive PC3 cells by elusing apoptosis. GLOI may be involved in prostate cancer progression, via the control of key molecules in the mitochondrial apoptotic mechanism, through NF-kB signaling pathway.
甘油醛-3-磷酸脱氢酶 I（GLOI）能解毒活性二羰基化合物，如甲基乙二醛（MG），MG 可直接或通过形成 MG 衍生加合物，作为生长抑制剂和凋亡诱导剂。GLOI 一直被认为是细胞增殖的一般标志物，但两者之间的直接联系尚未得到证实。本研究旨在阐明 GLOI 是否参与 LNCaP 和 PC3 人前列腺癌细胞的增殖控制，或者在这些细胞的生长调节中可能发挥不同的作用。
使用 RNA 干扰研究 GLOI 在细胞增殖或凋亡中的作用。通过[3H]胸苷掺入试验和流式细胞术评估细胞增殖，流式细胞术也用于分析凋亡。实时 TaqMan 聚合酶链反应和分光光度分析分别用于研究转录水平或特定活性。通过 Western blot 分析蛋白质水平。通过高效液相色谱法测量 MG。
我们发现 GLOI 不参与 LNCaP 和 PC3 细胞的增殖控制，但在侵袭性前列腺癌 PC3 细胞的凋亡中发挥作用，该作用通过涉及特定 MG 加合物和 NF-kB 信号通路的机制。
我们的数据首次系统地证明，GLOI 不能被视为细胞增殖的一般标志物，并且通过逃避凋亡，在侵袭性 PC3 细胞中作为一种生存促进因子发挥作用。GLOI 可能通过 NF-kB 信号通路，通过控制线粒体凋亡机制中的关键分子，参与前列腺癌的进展。
