Migfilin sensitizes cisplatin-induced apoptosis in human glioma cells in vitro
Migfilin sensitizes cisplatin-induced apoptosis in human glioma cells in vitro
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DOI：
10.1038/aps.2012.123
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发表时间：
2012-10-01
影响因子：
8.2
通讯作者：
Zhan, Qi-min
中科院分区：
文献类型：
--
作者：
Fan, Jing;Ou, Yun-wei;Zhan, Qi-min
目的：细丝蛋白结合LIM蛋白1，也称为migfilin，是一种骨架组织蛋白，在细胞-细胞外基质粘附时结合有丝分裂原诱导基因2。本研究的目的是探讨migfilin在顺铂诱导的人脑胶质瘤细胞凋亡中的作用，确定migfilin的功能结构域，并阐明migfilin调控顺铂相关化疗敏感性的分子机制。RNA干扰用于降低migfilin的表达。为了确定migfilin的功能结构域，用migfilin缺失突变体的质粒转染U-87 MG细胞。顺铂（40 μ mol/L）作用24 h后，MTS法检测细胞活力，DAPI染色法和TUNEL法检测细胞凋亡。结果：过表达migfilin可显著增强顺铂诱导的Hs 683、H4和U-87 MG细胞凋亡，而下调migfilin表达可抑制这些细胞株的化疗敏感性。单独的migfilin的N-末端区域能够增强顺铂诱导的细胞凋亡。然而，尽管存在的N-末端区域，三个LIM结构域的任何一个缺失的migfilin的突变体导致功能丧失。结论：migfilin的LIM 1 -3结构域通过调节凋亡相关蛋白，在胶质瘤细胞对顺铂诱导的凋亡敏感性中起重要作用。
Aim: Filamin binding LIM protein 1, also known as migfilin, is a skeleton organization protein that binds to mitogen-inducible gene 2 at cell-extracellular matrix adhesions. The aim of this study was to investigate the role of migfilin in cisplatin-induced apoptosis in human glioma cells, to determine the functional domains of migfilin, and to elucidate the molecular mechanisms underlying the regulation of cisplatin-related chemosensitivity.Methods: The human glioma cell lines Hs683, H4, and U-87 MG were transfected with pEGFP-C2-migfilin to elevate the expression level of migfilin. RNA interference was used to reduce the expression of migfilin. To determine the functional domains of migfilin, U-87 MG cells were transfected with plasmids of migfilin deletion mutants. After treatment with cisplatin (40 mu mol/L) for 24 h, the cell viability was assessed using the MTS assay, and the cell apoptotic was examined using the DAPI staining assay and TUNEL analysis. Expression levels of apoptosis-related proteins were detected by Western blot analysis.Results: Overexpression of migfilin significantly enhanced cisplatin-induced apoptosis in Hs683, H4, and U-87 MG cells, whereas down-regulation of migfilin expression inhibited the chemosensitivity of these cell lines. The N-terminal region of migfilin alone was able to enhance the cisplatin-induced apoptosis. However, despite the existence of the N-terminal region, mutants of migfilin with any one of three LIM domains deleted led to a function loss. Furthermore, apoptotic proteins (PARP and caspase-3) and the anti-apoptotic protein Bcl-xL were modulated by the expression level of migfilin in combination with cisplatin.Conclusion: The LIM1-3 domains of migfilin play a key role in sensitizing glioma cells to cisplatin-induced apoptosis through regulation of apoptosis-related proteins.