Department of Pediatric Surgery, Children's Hospital, Inselspital Bern, University of Bern, CH-3010 Bern, Switzerland.
Department of Human Genetics, Inselspital Bern, University of Bern, CH-3010 Bern, Switzerland.
Int J Mol Sci. 2024 Feb 14;25(4):2281. doi: 10.3390/ijms25042281.
Renal tumors comprise ~7% of all malignant pediatric tumors. Approximately 90% of pediatric kidney tumors comprise Wilms tumors, and the remaining 10% include clear cell sarcoma of the kidney, malignant rhabdoid tumor of the kidney, renal cell carcinoma and other rare renal tumors. Over the last 30 years, the role of cytokines and their receptors has been considerably investigated in both cancer progression and anti-cancer therapy. However, more effective immunotherapies require the cytokine profiling of each tumor type and comprehensive understanding of tumor biology. In this study, we aimed to investigate the activation of signaling pathways in response to cytokines in three pediatric kidney tumor cell lines, in WT-CLS1 and WT-3ab cells (both are Wilms tumors), and in G-401 cells (a rhabdoid kidney tumor, formerly classified as Wilms tumor). We observed that interferon-alpha (IFN-α) and interferon-gamma (IFN-γ) very strongly induced the activation of the STAT1 protein, whereas IL-6 and IFN-α activated STAT3 and IL-4 activated STAT6 in all examined tumor cell lines. STAT protein activation was examined by flow cytometry and Western blot using phospho-specific anti-STAT antibodies which recognize only activated (phosphorylated) STAT proteins. Nuclear translocation of phospho-STAT proteins upon activation with specific cytokines was furthermore confirmed by immunofluorescence. Our results also showed that both IFN-α and IFN-γ caused upregulation of major histocompatibility complex (MHC) class I proteins, however, these cytokines did not have any effect on the expression of MHC class II proteins. We also observed that pediatric kidney tumor cell lines exhibit the functional expression of an additional cytokine signaling pathway, the tumor necrosis factor (TNF)-α-mediated activation of nuclear factor kappa B (NF-κB). In summary, our data show that human pediatric renal tumor cell lines are responsive to stimulation with various human cytokines and could be used as in vitro models for profiling cytokine signaling pathways.
肾肿瘤约占所有儿童恶性肿瘤的 7%。大约 90%的儿童肾肿瘤由肾母细胞瘤组成，其余 10%包括肾透明细胞肉瘤、肾恶性横纹肌样瘤、肾细胞癌和其他罕见的肾肿瘤。在过去的 30 年中，细胞因子及其受体在癌症进展和抗癌治疗中的作用得到了广泛的研究。然而，更有效的免疫疗法需要对每种肿瘤类型进行细胞因子分析，并全面了解肿瘤生物学。在这项研究中，我们旨在研究三种儿科肾肿瘤细胞系对细胞因子的信号通路激活作用，这三种细胞系分别为 WT-CLS1 和 WT-3ab 细胞（均为肾母细胞瘤）和 G-401 细胞（横纹肌样肾肿瘤，以前归类为肾母细胞瘤）。我们观察到干扰素-α（IFN-α）和干扰素-γ（IFN-γ）非常强烈地诱导 STAT1 蛋白的激活，而 IL-6 和 IFN-α激活 STAT3，IL-4 激活 STAT6，所有检测的肿瘤细胞系均如此。STAT 蛋白的激活通过流式细胞术和 Western blot 进行检测，使用只识别激活（磷酸化）STAT 蛋白的磷酸化 STAT 抗体。通过用特异性细胞因子激活来进一步确认磷酸化 STAT 蛋白的核转位，通过免疫荧光实现。我们的结果还表明，IFN-α 和 IFN-γ 均可引起主要组织相容性复合体（MHC）I 类蛋白的上调，但这些细胞因子对 MHC II 类蛋白的表达没有任何影响。我们还观察到儿科肾肿瘤细胞系表现出另一种细胞因子信号通路的功能表达，即肿瘤坏死因子（TNF）-α介导的核因子 κB（NF-κB）的激活。总之，我们的数据表明，人儿科肾肿瘤细胞系对各种人细胞因子的刺激有反应，可以作为细胞因子信号通路分析的体外模型。
