de Souza Gustavo A, Godoy Lyris M F, Teixeira Veronica R, Otake Andreia H, Sabino Adão, Rosa José C, Dinarte Anemari R, Pinheiro Daniel G, Silva Wilson A, Eberlin Marcos N, Chammas Roger, Greene Lewis J
Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão - FAPESP, Brasil.
Proteomics. 2006 Mar;6(5):1460-70. doi: 10.1002/pmic.200500243.
Using 2-DE of total cell protein extracts, we compared soluble proteins from murine melanoma lines Tm1 and Tm5 with proteins from the nontumoral cell melan-a from which they were derived. Seventy-one of the 452 spots (average) detected with CBB were differentially accumulated, i.e., increased or decreased twofold. Forty-four spots were identified by PMF/MALDI-TOF, 15 with increased and 29 with decreased protein levels. SAGE showed that 17/34 (50%) of the differentially accumulated proteins, pI range 4-7, presented similar differences at the mRNA level. Major reductions in protein were observed in tumor cells of proteins that degrade reactive oxygen species (ROS). Decreases of > or = twofold in GST, superoxide dismutase, aldehyde dehydrogenase, thioredoxin, peroxiredoxin 2, and peroxiredoxin 6 protein were observed. SAGE indicated the reduction of other proteins involved in ROS degradation. As expected, the accumulation of exogenous peroxides was significantly higher in the tumor cells while the levels of glutathionylation were two times lower in the tumor cells compared to melan-a. The differential accumulation of proteins involved in oncogene/tumor suppressor pathways was observed. Melanoma cells can favor survival pathways activated by ROS by inhibiting p53 pathways and activation of Ras and c-myc pathways.
利用全细胞蛋白提取物的双向电泳技术，我们比较了小鼠黑色素瘤细胞系Tm1和Tm5中的可溶性蛋白与它们所源自的非肿瘤细胞黑色素-a中的蛋白。用考马斯亮蓝检测到的452个斑点（平均）中，有71个斑点存在差异积累，即增加或减少了两倍。通过肽质量指纹图谱/基质辅助激光解吸电离飞行时间质谱法鉴定出44个斑点，其中15个蛋白水平增加，29个蛋白水平降低。基因表达序列分析表明，17/34（50%）差异积累的蛋白，其等电点范围为4 - 7，在mRNA水平上呈现出类似的差异。在降解活性氧（ROS）的蛋白的肿瘤细胞中观察到蛋白的显著减少。谷胱甘肽S-转移酶、超氧化物歧化酶、醛脱氢酶、硫氧还蛋白、过氧化物酶2和过氧化物酶6蛋白减少了≥两倍。基因表达序列分析表明参与ROS降解的其他蛋白也减少。正如预期的那样，肿瘤细胞中外源过氧化物的积累显著更高，而与黑色素-a相比，肿瘤细胞中谷胱甘肽化水平低两倍。观察到参与癌基因/肿瘤抑制途径的蛋白的差异积累。黑色素瘤细胞可以通过抑制p53途径以及激活Ras和c-myc途径来促进由ROS激活的生存途径。
