Liu Xiaomeng, Liu Xiaosong, Zhao Lei, Wu Jianliang, Wang Xiaoliang, Hu Yuhua
Department of Neurosurgery, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei, China.
Department of Neurosurgery, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei, China.
Transl Oncol. 2025 Aug;58:102424. doi: 10.1016/j.tranon.2025.102424. Epub 2025 May 22.
Autophagy is an important factor in temozolomide (TMZ) resistance in glioblastoma (GBM). Receptor-interacting protein 2 (RIP2) is associated with autophagy, but its role and mechanism in regulating autophagy in GBM cells remain unclear. To analyze RIP2 expression in GBM in The Cancer Genome Atlas (TCGA) dataset. GBM cells were stimulated using recombinant human RIP2 protein (rRIP2) or RIP2 plasmid. Cell proliferation and apoptosis were assessed using CCK-8 assay and flow cytometry. Western blotting and immunofluorescence (IF) assays were performed to detect protein expression in cells and tumor tissues. Moreover, the relationship between RIP2-induced autophagy and TMZ resistance was verified in a GBM xenograft model. We determined that RIP2 expression was upregulated in GBM. rRIP2 and RIP2 overexpression induced TMZ resistance in the GBM cell lines. RIP2 overexpressing xenograft tumors have reduced sensitivity to TMZ. In addition, we showed that PD-L1 protein expression was upregulated in GBM tissues with RIP2 overexpression. rRIP2 and RIP2 overexpression induced autophagy in GBM cells through AMPK. Notably, RIP2 upregulated PD-L1 expression through the NF-κB signaling pathway, which induced autophagy and TMZ resistance in GBM cells. Moreover, NF-κB or autophagy inhibition reversed TMZ resistance in RIP2 overexpressing GBM cells in a xenograft model. In conclusion, RIP2 induces TMZ resistance in GBM cells by promoting autophagy through the NF-κB/PD-L1 signaling pathway, indicating that the RIP2/NF-κB/PD-L1 pathway may be therapeutic target for TMZ resistance.
自噬是胶质母细胞瘤（GBM）对替莫唑胺（TMZ）耐药的一个重要因素。受体相互作用蛋白2（RIP2）与自噬相关，但其在GBM细胞中调节自噬的作用和机制仍不清楚。为了分析癌症基因组图谱（TCGA）数据集中GBM中RIP2的表达情况。使用重组人RIP2蛋白（rRIP2）或RIP2质粒刺激GBM细胞。采用CCK-8法和流式细胞术评估细胞增殖和凋亡。进行蛋白质印迹和免疫荧光（IF）分析以检测细胞和肿瘤组织中的蛋白质表达。此外，在GBM异种移植模型中验证了RIP2诱导的自噬与TMZ耐药之间的关系。我们确定GBM中RIP2表达上调。rRIP2和RIP2过表达诱导GBM细胞系对TMZ耐药。RIP2过表达的异种移植肿瘤对TMZ的敏感性降低。此外，我们发现RIP2过表达的GBM组织中PD-L1蛋白表达上调。rRIP2和RIP2过表达通过AMPK诱导GBM细胞自噬。值得注意的是，RIP2通过NF-κB信号通路上调PD-L1表达，从而诱导GBM细胞自噬和TMZ耐药。此外，在异种移植模型中，NF-κB或自噬抑制可逆转RIP2过表达的GBM细胞中的TMZ耐药。总之，RIP2通过NF-κB/PD-L1信号通路促进自噬，从而诱导GBM细胞对TMZ耐药，表明RIP2/NF-κB/PD-L1通路可能是TMZ耐药的治疗靶点。
