Kahlos K, Anttila S, Asikainen T, Kinnula K, Raivio K O, Mattson K, Linnainmaa K, Kinnula V L
Department of Internal Medicine, University of Oulu, Oulu, Finland.
Am J Respir Cell Mol Biol. 1998 Apr;18(4):570-80. doi: 10.1165/ajrcmb.18.4.2943.
We hypothesized that manganese superoxide dismutase (MnSOD), known to be induced in rat mesothelial cells by asbestos fibers, cytokines, and hyperoxia, may also be induced in asbestos-related pleural diseases such as mesothelioma. MnSOD was assessed in healthy human pleural mesothelium (n = 6), in biopsy samples of human pleural mesothelioma (n = 7), in transformed nonmalignant human mesothelial cells (Met5A), and in two human mesothelioma cell lines (M14K and M38K) established from the tumor tissue of mesothelioma patients. There was no MnSOD immunoreactivity in five of the six samples of healthy pleural mesothelium, whereas MnSOD immunoreactivity was high in the tumor cells in all the mesothelioma samples. Northern blotting, immunohistochemistry, Western blotting, and specific activity measurements showed lower MnSOD in the nonmalignant Met5A mesothelial cells than in the M14K and M38K mesothelioma cells. In additional experiments the mesothelial and mesothelioma cells were exposed to menadione, which generates superoxide intracellularly, and to epirubicin, a cytotoxic drug commonly used to treat mesothelioma. The M38K mesothelioma cells were most resistant to menadione and epirubicin when assessed by LDH release or by adenine nucleotide (ATP, ADP, and AMP) depletion. These same cells showed not only the highest MnSOD levels, but also the highest mRNA levels and activities of catalase, whereas glutathione peroxidase and glutathione reductase levels did not differ significantly. We conclude that MnSOD expression is low in healthy human pleural mesothelium and high in human malignant mesothelioma. The most resistant mesothelioma cells contained coordinated induction of MnSOD and catalase.
我们推测,已知在大鼠间皮细胞中可被石棉纤维、细胞因子和高氧诱导产生的锰超氧化物歧化酶(MnSOD),在诸如间皮瘤等石棉相关胸膜疾病中也可能被诱导产生。我们对健康人胸膜间皮(n = 6)、人胸膜间皮瘤活检样本(n = 7)、转化的非恶性人间皮细胞(Met5A)以及从间皮瘤患者肿瘤组织建立的两个人间皮瘤细胞系(M14K和M38K)中的MnSOD进行了评估。在六个健康胸膜间皮样本中的五个中未检测到MnSOD免疫反应性,而在所有间皮瘤样本的肿瘤细胞中MnSOD免疫反应性较高。Northern印迹法、免疫组织化学、Western印迹法和比活性测量显示,非恶性Met5A间皮细胞中的MnSOD低于M14K和M38K间皮瘤细胞。在额外的实验中,将间皮细胞和间皮瘤细胞暴露于能在细胞内产生超氧化物的甲萘醌以及常用于治疗间皮瘤的细胞毒性药物表柔比星。通过乳酸脱氢酶释放或腺嘌呤核苷酸(ATP、ADP和AMP)消耗评估时,M38K间皮瘤细胞对甲萘醌和表柔比星的耐受性最强。这些相同的细胞不仅显示出最高的MnSOD水平,还显示出过氧化氢酶的最高mRNA水平和活性,而谷胱甘肽过氧化物酶和谷胱甘肽还原酶水平没有显著差异。我们得出结论,MnSOD在健康人胸膜间皮中的表达较低,而在人恶性间皮瘤中较高。耐受性最强的间皮瘤细胞中MnSOD和过氧化氢酶协同诱导。