Department of Neurosurgery, Chonnam National University Hwasun Hospital, Jeollanam-do, Republic of Korea.
Anticancer Res. 2013 Jun;33(6):2525-33.
The aim of this study was the immunological characterization of glioblastoma cells. Glioblastoma cell lines were cultured in serum and serum-free neurobasal (NBE) medium conditions. These cell lines were characterized by flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), western blot and natural killer (NK) cell-cytotoxicity assays. A previously described NK cell expansion method that uses K562 cells expressing interleukin (IL)-15 and 4-1 BB Ligand (BBL) (K562-mb15-41BBL) was used. RT-PCR and western blots for the expression of tumor-associated antigens (TAAs), were carried out in 32 glioblastoma and seven normal brain tissues. U87 and U343 tumor cell lines showed increased expression for major histocompatibility complex (MHC)-I and -II molecules. No significant differences in the levels of CD133, MHC class I/II, MHC class I-related chain A (MICA), MICB, UL16 binding protein 1-3 (ULBP 1-3) expression in these cell lines and in NK cell cytotoxicity were observed between serum and NBE conditions. Regardless of culture conditions, U87 and U343 cell lines were sensitive to expanded NK cells, with median cytotoxicities at 4:1 effector/target ratio of 43.2% and 46.5%, respectively. In RT-PCR, U343 and U87 showed the expression of most TAAs at a high ratio compared with U251. Western blots demonstrated positive expression for BIRC5, CD99 and ERBB2 in U251, U87 and U343 cell lines and tissues. These highly-expressed TAAs such as BIRC5, CD99 and ERBB2 in glioblastoma tissue could be the targets for immunotherapy. U87 and U343 cell lines could be useful for studying the efficacy of immunotherapy related to various TAAs and NK cell immunotherapy.
本研究旨在对胶质母细胞瘤细胞进行免疫学特征分析。将胶质母细胞瘤细胞系在含血清和无血清神经基底（NBE）培养基的条件下进行培养。通过流式细胞术、逆转录-聚合酶链反应（RT-PCR）、western blot 和自然杀伤（NK）细胞细胞毒性测定对这些细胞系进行了特征描述。使用先前描述的使用表达白细胞介素（IL）-15 和 4-1BB 配体（BBL）的 K562 细胞（K562-mb15-41BBL）的 NK 细胞扩增方法。对 32 例胶质母细胞瘤和 7 例正常脑组织进行了肿瘤相关抗原（TAA）表达的 RT-PCR 和 western blot。U87 和 U343 肿瘤细胞系显示主要组织相容性复合体（MHC）-I 和 -II 分子表达增加。在这些细胞系和 NK 细胞细胞毒性中，血清和 NBE 条件下 CD133、MHC Ⅰ/Ⅱ、MHC Ⅰ类相关链 A（MICA）、MICB、UL16 结合蛋白 1-3（ULBP 1-3）表达水平无显著差异。无论培养条件如何，U87 和 U343 细胞系对扩增的 NK 细胞均敏感，在 4:1 效靶比时的中位细胞毒性分别为 43.2%和 46.5%。在 RT-PCR 中，U343 和 U87 与 U251 相比，大多数 TAA 的表达比例较高。western blot 显示 U251、U87 和 U343 细胞系和组织中 BIRC5、CD99 和 ERBB2 的阳性表达。胶质母细胞瘤组织中这些高表达的 TAA，如 BIRC5、CD99 和 ERBB2，可能是免疫治疗的靶点。U87 和 U343 细胞系可用于研究与各种 TAA 和 NK 细胞免疫治疗相关的免疫治疗疗效。
