Induction of autophagy in temozolomide treated malignant gliomas
Induction of autophagy in temozolomide treated malignant gliomas
复制标题
DOI：
10.1111/j.1440-1789.2010.01197.x
复制
发表时间：
2011-10-01
期刊：
影响因子：
2.3
通讯作者：
Fujii, Yukihiko
中科院分区：
文献类型：
--
作者：
Natsumeda, Manabu;Aoki, Hiroshi;Fujii, Yukihiko
自噬是蛋白质降解的动态过程。在神经胶质瘤细胞系中已经注意到替莫唑胺（TMZ）诱导自噬。28个标本，从14例TMZ治疗前后，进行了分析，以探讨是否诱导自噬可以检测到手术标本的免疫组化分析。采用抗轻链3亚型B（LC 3 B）和抗溶酶体相关膜蛋白1（LAMP 1）抗体通过免疫组织化学分析监测大细胞自噬;通过抗LAMP 2A抗体免疫染色监测分子伴侣介导的自噬。此外，LC 3B蛋白的Western印迹检测从3例患者的冷冻保存组织中获得的6个标本。所有标本均显示胶质瘤细胞胞质中每种免疫染色呈点状染色，表明诱导了自噬。LC 3B、LAMP 1和LAMP 2A免疫染色半定量评分为1 - 3分。与治疗前评分（5.2 +/-1.5）相比，TMZ治疗后的三项评分（6.4 +/- 1.2）显着增加（P = 0.020）。LC 3B的蛋白质印迹显示TMZ处理后LC 3B-I和LC 3B-II表达增加。本研究证明了用免疫组化染色法监测外科手术标本的自噬是可行的。这些结果表明TMZ诱导自噬。
Autophagy is a dynamic process of protein degradation. Induction of autophagy by temozolomide (TMZ) has been noted in glioma cell lines. Twenty-eight specimens, obtained from 14 patients before and after TMZ treatment, were analyzed to investigate whether induction of autophagy could be detected in surgical specimens by immunohistochemical analysis. Macroautophagy was monitored by immunohistochemical analysis employing anti-light chain 3 isoform B (LC3B) and anti-lysosome-associated membrane protein 1 (LAMP1) antibodies; chaperone-mediated autophagy was monitored by anti-LAMP2A antibody immunostaining. Furthermore, detection of LC3B protein by Western blotting was performed on six specimens obtained from the preserved frozen tissues of three patients. All specimens showed dot-like staining for each immunostain in the cytoplasm of glioma cells, indicating induction of autophagy. LC3B, LAMP1 and LAMP2A immunostains were semiquantitatively scored from 1 to 3 points. Combination of the three scores after TMZ treatment (6.4 +/- 1.2) showed a significant increase (P = 0.020) compared to pre-treatment scores (5.2 +/- 1.5). Western blotting for LC3B showed increased LC3B-I and LC3B-II expression after TMZ treatment. The present study proved that autophagy monitoring by immunohistochemical staining of surgical specimens was feasible. These results suggest that autophagy is induced by TMZ.