Jung Yieun, Ahn So-Hee, Park Hyunju, Park Sang Hui, Choi Kyungsun, Choi Chulhee, Kang Jihee Lee, Choi Youn-Hee
Department of physiology, Ewha Womans University School of medicine, Seoul, Republic of Korea.
Tissue Injury Defense Research Center, Ewha Womans University School of medicine, Seoul, Republic of Korea.
Cell Physiol Biochem. 2018;48(3):1332-1346. doi: 10.1159/000492092. Epub 2018 Jul 26.
BACKGROUND/AIMS: Glioblastoma multiforme (GBM) is the most common primary brain tumor in adults. The defining characteristics of GBM are diffuse infiltration of tumor cells into normal brain parenchyma, rapid growth, a high degree of infiltration of microglia and macrophages, and the presence of necrosis. Microglia/macrophages are frequently found in gliomas and they extensively infiltrate GBM tissue, up to 30% of total tumor mass. However, little is known about the effect of necrotic cells (NCs) on microglia infiltration in GBM and the tumor-infiltrating microglia-induced factors in GBMs.
In this study, to address whether necrosis or necrosis-exposed GBM cells affect the degree of microglia/macrophage infiltration, migration and invasion/infiltration assays were performed. Culture supernatants and nuclear extracts of CRT-MG cells treated or untreated with necrotic cells were analyzed using a chemokine array and electrophoretic mobility shift assay, respectively.
The presence of NCs promoted the migration/infiltration of microglia, and GBM cell line CRT-MG cells exposed to NCs further enhanced the migration and infiltration of HMO6 microglial cells. Treatment with NCs induced mRNA and protein expression of chemokines such as
These results provide evidence that NCs induce the expression of CCL2/MCP-1 and CCL20/MIP-3α in glioblastoma cells through activation of NF-κB and AP-1 and facilitate the infiltration of microglia into tumor tissues.
背景/目的：多形性胶质母细胞瘤（GBM）是成人中最常见的原发性脑肿瘤。GBM的定义特征包括肿瘤细胞向正常脑实质的弥漫性浸润、快速生长、小胶质细胞和巨噬细胞的高度浸润以及坏死的存在。小胶质细胞/巨噬细胞经常在胶质瘤中发现，并且它们广泛浸润GBM组织，占肿瘤总质量的30%。然而，关于坏死细胞（NCs）对GBM中小胶质细胞浸润的影响以及GBM中肿瘤浸润小胶质细胞诱导因子知之甚少。
在本研究中，为了探讨坏死或暴露于坏死的GBM细胞是否影响小胶质细胞/巨噬细胞浸润程度，进行了迁移和侵袭/浸润实验。分别使用趋化因子阵列和电泳迁移率变动分析，对用坏死细胞处理或未处理的CRT-MG细胞的培养上清液和核提取物进行分析。
NCs的存在促进了小胶质细胞的迁移/浸润，并且暴露于NCs的GBM细胞系CRT-MG细胞进一步增强了HMO6小胶质细胞的迁移和浸润。用NCs处理诱导了CRT-MG细胞中趋化因子如单核细胞趋化蛋白-1（CCL2/MCP-1）和巨噬细胞炎性蛋白-3α（CCL20/MIP-3α）的mRNA和蛋白表达。特别是，在经NC处理的CRT-MG细胞中CCL2/MCP-1和CCL20/MIP-3α显著增加。NCs诱导转录因子核因子（NF）-κB和激活蛋白1（AP-1）与CCL2/MCP-1和CCL20/MIP-3α启动子的DNA结合，导致CRT-MG细胞中CCL2/MCP-1和CCL20/MIP-3α的mRNA和蛋白表达增加。
这些结果提供了证据，表明NCs通过激活NF-κB和AP-1诱导胶质母细胞瘤细胞中CCL2/MCP-1和CCL20/MIP-3α的表达，并促进小胶质细胞向肿瘤组织的浸润。
