Milicevic Zorka, Kasapovic Jelena, Gavrilovic Ljubica, Milovanovic Zorka, Bajic Vladan, Spremo-Potparevic Biljana
Laboratory of Molecular Biology and Endocrinology, "Vinca" Institute of Nuclear Sciences, Mihaila Petrovica Alasa 12-14, University of Belgrade, 11 001 Belgrade, Serbia.
National Center of Cancer Research (NCRC), Institute for Oncology and Radiology of Serbia, Pasterova 14, 11 000 Belgrade, Serbia.
EXCLI J. 2014 Jun 23;13:691-708. eCollection 2014.
It is well recognized that cancers develop and grow as a result of disordered function of tumor suppressor genes and oncogenes, which may be exploited for screening purposes. Extensive evidence indicated tumor suppressor protein p53 as candidate marker for mutation identification. We have investigated mutant p53 protein expression in human breast tumors in relation to antioxidant status deficiency. The study included 100 breast cancer patients. p53 protein expression was evaluated by Western blot assay and immunostaining using a CM-1, DO-7 and Pab240 antibodies. Antioxidant parameters and lipid peroxidation were estimated by biochemical analyses. Western blotting with epitopespecific monoclonal antibody Pab240 strongly suggests that nuclear extracts from breast cancer cells express mutant forms of p53. It is of interest that the mutant forms of p53 overexpression in conjunction with the appearance of nuclear bodies are observed in highly aggressive carcinomas. Expression of isoform Δp53 (45 kDa) and isoform of ~ 29 kDa were more common in cases with LN metastasis. These studies point out the molecular consequences of oxidative stress (lipid peroxides, LP, p<0.001) and antioxidant status deficiency (copper, zinc superoxid dismutase, SOD, p<0.001; catalase, CAT, p<0.01; glutathione reductase, GR, p<0.001; glutathione, GSH, p<0.05) and indicate the importance of p53 mutation as the commonest genetic alteration detected in breast cancer cells. The expression of mutant p53 is correlated to increased lipid peroxides (0.346, p<0.05 ) and lowered antioxidant activity of CAT (- 0.437, p<0.01) in the breast cancer patients.
众所周知,癌症的发生和发展是由于肿瘤抑制基因和癌基因功能紊乱所致,这可用于筛查目的。大量证据表明肿瘤抑制蛋白p53是用于识别突变的候选标志物。我们研究了人类乳腺肿瘤中突变型p53蛋白表达与抗氧化状态缺乏之间的关系。该研究纳入了100名乳腺癌患者。通过蛋白质免疫印迹分析以及使用CM-1、DO-7和Pab240抗体进行免疫染色来评估p53蛋白表达。通过生化分析评估抗氧化参数和脂质过氧化。用表位特异性单克隆抗体Pab240进行的蛋白质免疫印迹强烈表明,乳腺癌细胞核提取物表达p53的突变形式。有趣的是,在高侵袭性癌中观察到p53突变形式的过表达与核体的出现有关。在有淋巴结转移的病例中,异构体Δp53(45 kDa)和~29 kDa异构体的表达更为常见。这些研究指出了氧化应激(脂质过氧化物,LP,p<0.001)和抗氧化状态缺乏(铜、锌超氧化物歧化酶,SOD,p<0.001;过氧化氢酶,CAT,p<0.01;谷胱甘肽还原酶,GR,p<0.001;谷胱甘肽,GSH,p<0.05)的分子后果,并表明p53突变作为在乳腺癌细胞中检测到的最常见基因改变的重要性。在乳腺癌患者中,突变型p53的表达与脂质过氧化物增加(0.346,p<0.05)和CAT抗氧化活性降低(-0.437,p<0.01)相关。