The positive regulatory loop of TCF4N/p65 promotes glioblastoma tumourigenesis and chemosensitivity.
The positive regulatory loop of TCF4N/p65 promotes glioblastoma tumourigenesis and chemosensitivity.
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TCF4N/p65 的正调控环促进胶质母细胞瘤肿瘤发生和化疗敏感性
DOI：
10.1002/ctm2.1042
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发表时间：
2022-09
影响因子：
10.6
通讯作者：
Zou, Jian
中科院分区：
文献类型：
--
作者：
Hu, Yaling;Zhang, Bo;Lu, Peihua;Wang, Jingying;Chen, Cheng;Yin, Ying;Wan, Quan;Wang, Jingjing;Jiao, Jiantong;Fang, Xiangming;Pu, Zhening;Gong, Lingli;Ji, Li;Zhu, Lingpeng;Zhang, Rui;Zhang, Jia;Yang, Xusheng;Wang, Qing;Huang, Zhaohui;Zou, Jian
NF-κB 信号传导与癌症的发病机制和治疗耐药性广泛相关。由于其在细胞衰老和衰老相关分泌表型（SASP）中的关键功能，其抗癌作用越来越受到关注。因此，在应用 NF-κB 抑制剂之前，有必要深入了解 NF-κB 在癌症中的功能和调控。我们建立了表达异位TCF4N（β-连环蛋白相互作用转录因子TCF7L2的同种型）的胶质母细胞瘤（GBM）细胞系，并在体外和体内评估了其在GBM肿瘤发生和化疗中的功能。在 p65 敲除或磷酸化模拟 (S536D) 细胞系中，通过体外和体内功能实验研究了 TCF4N 和 NF-κB 信号在促进肿瘤发生和化疗敏感性中的双重作用和相关性。进行RNA测序和计算分析、免疫沉淀和泛素化测定、小基因剪接测定和荧光素酶报告基因测定，以确定TCF4N和NF-κB的p65亚基之间正反馈调节环的潜在机制。使用靶向 TCF4N（4N）的真核细胞穿透肽来证实其治疗意义。我们的结果表明，p65 亚基 Ser 536 (S536) 磷酸化和核积累是 GBM 的一个有前途的预后标志物，并赋予 NF-κB 促进肿瘤发生和化疗敏感性的双重功能。 GBM 中 p65 S536 磷酸化和核稳定性受 TCF4N 调节。 TCF4N 结合 p65，诱导 p65 磷酸化和核转位，抑制其泛素化/降解，随后促进 NF-κB 活性。 p65 S536 磷酸化对于 TCF4N 主导的不依赖于衰老的 SASP、GBM 肿瘤发生、肿瘤干细胞样细胞分化和化疗敏感性至关重要。 p65 的激活与 TCF4N 的可变剪接密切相关，这可能是 GBM 中 TCF4N 和 p65 之间的正反馈调节环。 4N 增加了化学敏感性，凸显了一种新的抗癌策略。我们的研究通过与 p65 的相互调节，明确了 TCF4N 作为 NF-κB 新型调节剂的关键作用，并为 GBM 抑制提供了新途径。 TCF4N/p65 的正调节环促进胶质母细胞瘤 (GBM) 肿瘤发生和化疗敏感性：TCF4N 通过结合和促进 p65 S536 磷酸化、核转位和稳定性来上调 NF-κB 活性； p65 促进 TCF7L2 前 mRNA 的选择性剪接，导致 TCF4N 增加。
NF‐κB signaling is widely linked to the pathogenesis and treatment resistance in cancers. Increasing attention has been paid to its anti‐oncogenic roles, due to its key functions in cellular senescence and the senescence‐associated secretory phenotype (SASP). Therefore, thoroughly understanding the function and regulation of NF‐κB in cancers is necessary prior to the application of NF‐κB inhibitors. We established glioblastoma (GBM) cell lines expressing ectopic TCF4N, an isoform of the β‐catenin interacting transcription factor TCF7L2, and evaluated its functions in GBM tumorigenesis and chemotherapy in vitro and in vivo. In p65 knock‐out or phosphorylation mimic (S536D) cell lines, the dual role and correlation of TCF4N and NF‐κB signaling in promoting tumorigenesis and chemosensitivity was investigated by in vitro and in vivo functional experiments. RNA‐seq and computational analysis, immunoprecipitation and ubiquitination assay, minigene splicing assay and luciferase reporter assay were performed to identify the underlying mechanism of positive feedback regulation loop between TCF4N and the p65 subunit of NF‐κB. A eukaryotic cell‐penetrating peptide targeting TCF4N, 4N, was used to confirm the therapeutic significance. Our results indicated that p65 subunit phosphorylation at Ser 536 (S536) and nuclear accumulation was a promising prognostic marker for GBM, and endowed the dual functions of NF‐κB in promoting tumorigenesis and chemosensitivity. p65 S536 phosphorylation and nuclear stability in GBM was regulated by TCF4N. TCF4N bound p65, induced p65 phosphorylation and nuclear translocation, inhibited its ubiquitination/degradation, and subsequently promoted NF‐κB activity. p65 S536 phosphorylation was essential for TCF4N‐led senescence‐independent SASP, GBM tumorigenesis, tumor stem‐like cell differentiation and chemosensitivity. Activation of p65 was closely connected to alterative splicing of TCF4N, a likely positive feedback regulation loop between TCF4N and p65 in GBM. 4N increased chemosensitivity, highlighting a novel anti‐cancer strategy. Our study defined key roles of TCF4N as a novel regulator of NF‐κB through mutual regulation with p65 and provided a new avenue for GBM inhibition. The positive regulatory loop of TCF4N/p65 promotes glioblastoma (GBM) tumourigenesis and chemosensitivity: TCF4N upregulates NF‐κB activity by binding and promoting p65 S536 phosphorylation, nuclear‐translocation and stability; p65 promotes alternative splicing of TCF7L2 pre‐mRNA, resulting in an increase of TCF4N.
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DOI：
10.1146/annurev-pathol-121808-102144
发表时间：
2010
期刊：
Annual review of pathology
影响因子：
--
作者：
Coppé JP;Desprez PY;Krtolica A;Campisi J
通讯作者：
Campisi J
影响因子：
10.5
作者：
Chien, Yuchen;Scuoppo, Claudio;Lowe, Scott W.
通讯作者：
Lowe, Scott W.
影响因子：
3.8
作者：
Jing H;Lee S
通讯作者：
Lee S
影响因子：
10.5
作者：
Birch J;Gil J
通讯作者：
Gil J
影响因子：
4.6
作者：
Hu, J;Haseebuddin, M;Colburn, NH
通讯作者：
Colburn, NH