Fukuda Mieko E, Iwadate Yasuo, Machida Toshio, Hiwasa Takaki, Nimura Yoshinori, Nagai Yuichiro, Takiguchi Masaki, Tanzawa Hideki, Yamaura Akira, Seki Naohiko
Department of Neurological Surgery, Chiba University Graduate School of Medicine, Chiba, Japan.
Cancer Res. 2005 Jun 15;65(12):5190-4. doi: 10.1158/0008-5472.CAN-04-4134.
Cathepsin D is an aspartyl protease involved in protein catabolism and tissue remodeling which can be secreted from cancer cells. To identify a potential serum marker for gliomas, we investigated the gene expression levels of cathepsin D in 87 tissue samples and measured the protein concentrations in sera of glioma patients. The tissue samples consisted of 43 glioblastomas, 13 anaplastic astrocytomas, 22 astrocytomas, and 9 normal brain tissues. The results of real-time quantitative reverse transcription-PCR analysis showed that cathepsin D transcript levels became significantly higher as the glioma grade advanced (P = 0.0466, glioblastoma and anaplastic astrocytoma; P = 0.0008, glioblastoma and astrocytoma; P = 0.0271, glioblastoma and normal brain tissue; unpaired t test). Immunohistochemical analysis with anti-cathepsin D antibody revealed dense and spotty staining in the tumor cells with high transcript levels. The low expression of cathepsin D significantly correlated with long survival of the glioma patients. Furthermore, the glioblastoma patients with high gene expression of cathepsin D lived significantly shorter than those with low expression (P = 0.0104, Cox-Mantel log-rank test) and frequently had leptomeningeal dissemination (P = 0.0016, chi2 test). The multivariate analysis confirmed that the cathepsin D expression level was an independent predictor for short survival (P = 0.0102, Cox proportional hazard regression model). Measurement of the serum cathepsin D concentrations by ELISA showed a significant increase in the patients with high-grade gliomas as compared with the low-grade tumors (P = 0.0081, chi2 test). These results collectively suggest that cathepsin D could be a potential serum marker for the prediction of aggressive nature of human gliomas.
组织蛋白酶D是一种参与蛋白质分解代谢和组织重塑的天冬氨酸蛋白酶,可由癌细胞分泌。为了鉴定一种潜在的胶质瘤血清标志物,我们研究了87份组织样本中组织蛋白酶D的基因表达水平,并测量了胶质瘤患者血清中的蛋白质浓度。组织样本包括43例胶质母细胞瘤、13例间变性星形细胞瘤、22例星形细胞瘤和9例正常脑组织。实时定量逆转录 - PCR分析结果显示,随着胶质瘤分级的升高,组织蛋白酶D转录水平显著升高(胶质母细胞瘤与间变性星形细胞瘤比较,P = 0.0466;胶质母细胞瘤与星形细胞瘤比较,P = 0.0008;胶质母细胞瘤与正常脑组织比较,P = 0.0271;未配对t检验)。用抗组织蛋白酶D抗体进行免疫组织化学分析显示,转录水平高的肿瘤细胞中有密集和斑点状染色。组织蛋白酶D的低表达与胶质瘤患者的长期生存显著相关。此外,组织蛋白酶D基因表达高的胶质母细胞瘤患者的生存期明显短于低表达患者(P = 0.0104,Cox - Mantel对数秩检验),并且经常发生软脑膜播散(P = 0.0016,卡方检验)。多变量分析证实,组织蛋白酶D表达水平是短期生存的独立预测指标(P = 0.0102,Cox比例风险回归模型)。通过ELISA测量血清组织蛋白酶D浓度显示,与低级别肿瘤相比,高级别胶质瘤患者的血清组织蛋白酶D浓度显著升高(P = 0.0081,卡方检验)。这些结果共同表明,组织蛋白酶D可能是预测人类胶质瘤侵袭性的潜在血清标志物。