Department of Translational Neuroscience, University Medical Center Utrecht (UMCU) Brain Center, Utrecht University, 3584 CX Utrecht, The Netherlands.
Department of Human Genetics, University of Liège, 4000 Liège, Belgium.
Cells. 2024 Nov 14;13(22):1883. doi: 10.3390/cells13221883.
Previous works have shown that the expression of Class-II-Transactivator (CIITA) in tumor cells reduces the growth of glioblastoma (GB) in animal models, but immune effects cannot solely explain this. Here, we searched for immune-independent effects of CIITA on the proliferation of GB.
Murine GL261 and human U87, GM2 and GM3 malignant glioma cells were transfected with CIITA. NSG (immunodeficient) and nude (athymic) mice were injected in the striatum with GL261-wildtype (-WT) and -CIITA, and tumor growth was assessed by immunohistology and luminescence reporter genes. Clonogenic, sphere-formation, and 3D Matrigel-based in vitro growth assays were performed to compare the growth of WT versus CIITA-expressing murine and human cells. Bulk RNA sequencing and RT qRT-PCR profiler arrays were performed on these four cell lines to assess RNA expression changes following CIITA transfection. Western blot analysis on several proliferation-associated proteins was performed.
The intracerebral growth of murine GL261-CIITA cells was drastically reduced both in immunodeficient and athymic mice. Tumor growth was reduced in vitro in three of the four cell types. RNA sequencing and RT profiler array experiments revealed a modulation of gene expression in the PI3-Akt, MAPK- and cell-cycle regulation pathways following CIITA overexpression. Western blot analysis showed an upregulation of p27 in the growth-inhibited cells following this treatment. PDGFR-beta was downregulated in all cells. We did not find consistent regulation of other proteins involved in GB proliferation.
Proliferation is drastically reduced by CIITA in GB, both in vivo and in vitro, notably in association with p27-mediated inhibition of cell-cycle pathways.
先前的研究表明，肿瘤细胞中 II 类转录激活因子（CIITA）的表达可降低动物模型中胶质母细胞瘤（GB）的生长，但免疫作用并不能完全解释这一现象。在此，我们研究了 CIITA 对 GB 增殖的免疫独立影响。
用 CIITA 转染小鼠 GL261 和人 U87、GM2 和 GM3 恶性神经胶质瘤细胞。将 NSG（免疫缺陷）和裸鼠（无胸腺）脑纹状体注射 GL261-野生型（-WT）和 -CIITA，通过免疫组化和发光报告基因评估肿瘤生长。进行集落形成、球体形成和 3D Matrigel 体外生长实验，比较 WT 与 CIITA 表达的小鼠和人细胞的生长情况。对这四种细胞系进行批量 RNA 测序和 RT qRT-PCR 分析，以评估 CIITA 转染后 RNA 表达的变化。对几种与增殖相关的蛋白进行 Western blot 分析。
CIITA 转染的 GL261 细胞在免疫缺陷和无胸腺小鼠中的颅内生长明显减少。在这四种细胞类型中的三种中，体外肿瘤生长均受到抑制。RNA 测序和 RT 分析表明，CIITA 过表达后，PI3-Akt、MAPK 和细胞周期调控通路的基因表达发生了变化。Western blot 分析表明，经该治疗后，生长受到抑制的细胞中 p27 上调。所有细胞中的 PDGFR-beta 下调。我们没有发现其他与 GB 增殖相关的蛋白存在一致的调节作用。
CIITA 可明显抑制 GB 的体内外增殖，尤其是与 p27 介导的细胞周期途径抑制有关。
