NF-κB /UCHL5/TGF-βR1介导辐射诱导的糖代谢重编程促进宫颈癌加速重建

The mechanisms underlying “accelerated repopulation” in cervical cancer after radiotherapy remain to be elucidated. Radiation-induced metabolic reprogramming plays important roles in tumorigenesis and progression. Our study showed that UCHL5 was up-regulated in radioresistant cervical cancer cells in a NF-κB-dependent manner. Overexpression of UCHL5 promoted glycolysis and increased TGF-βR1 expression in cervical cancer cells. We further demonstrated that UCHL5 expression level in cervical cancer tissues was significantly higher than that in normal cervical tissues. Based on these data , we hypothesized that NF-κB/UCHL5/TGF-βR1 mediated radiation-induced glucose metabolic reprogramming promotes accelerated repopulation in cervical cancer. In the current study, reporter gene assay, EMSA and Chip assay will be employed to uncover how NFKB regulate UCHL5 expression. Gain-of-function and loss-of-function will be used to verify TGF-βR1 is a functional target of UCHL1. Glucose metabolic reprogramming assay will be performed to observe the effects of NF-κB/UCHL5/TGF-βR1 on accelerated repopulation of cervical cancer. The study will deepen our understanding of tumor metabolic reprogramming and help us to find novel therapeutic targets for tumor accelerated repopulation.

肿瘤放疗后残存癌细胞“加速重建”的调控机制仍未完全阐明。辐射诱导的代谢重编程是放疗后残癌加速重建的重要基础。我们的前期工作发现，UCHL5在宫颈癌组织中以及放疗后残留Hela细胞中表达明显升高。辐射诱导UCHL5在宫颈癌细胞表达，显著促进宫颈癌细胞糖酵解。NF-κB介导了辐射对UCHL5的表达诱导作用，辐射和UCHL5可以促进宫颈癌细胞TGF-βR1的表达。提示：NF-κB /UCHL5 /TGF-βR1介导辐射诱导的糖代谢重编程促进宫颈癌加速重建。本项目将借助报告基因实验、EMSA、Chip明确NFKB调控UCHL5表达的机制，通过干预和拮抗实验确证TGF-βR1是UCHL5的功能靶分子，利用体内外糖代谢重编程实验最终验证“NF-κB /UCHL5/TGF-βR1” 这一信号通路促进宫颈癌放疗后残癌加速重建”这一理论假设，为深入理解放疗后宫颈癌残癌加速重建的调控机制提供理论依据。

去泛素化酶的异常表达影响肿瘤的发生发展、细胞侵袭转移和放化疗抵抗等多个方面。我们的前期工作发现，去泛素化酶UCHL5在放疗后残留宫颈癌细胞中表达明显升高，但其相关调控机制未明。. 本项目主要研究结果：1. UCHL5在宫颈癌组织中高表达并与患者预后负相关。2. NF-κB 对UCHL5 表达具有正向调节作用，而辐射和UCHL5可以促进宫颈癌细胞TGF-βR1的表达。3. 辐射诱导UCHL5在宫颈癌细胞表达，显著促进宫颈癌细胞糖酵解。4. UCHL5降低了宫颈癌细胞的辐射敏感性，增强细胞增殖活性和裸鼠移植瘤形成能力。5. UCHL5可有效增强宫颈癌Hela细胞的凋亡抵抗能力，加速细胞周期进展，促进细胞迁移，缓解辐照引起的细胞G2/M期阻滞。6. 转录组测序和串联亲和纯化结果显示，UCHL5和XRCC6相互作用。7. 过表达UCHL5可降低XRCC6的泛素化水平、提高其蛋白质表达水平，而敲减UCHL5可以降低XRCC6的表达水平，并且，敲减XRCC6可以抑制UCHL5对宫颈癌细胞促增殖效应。8. XRCC6在宫颈癌组织的表达水平增高并且和UCHL5的表达呈正相关。9. UCHL5可通过去泛素化酶活性增强β-catenin的稳定性，促进宫颈癌细胞Wnt/β-catenin信号通路下游效应分子TCF1/TCF7、CD44、c-Myc和c-Jun的表达。. 上述结果提示，辐射和NF-κB可以诱导UCHL5在宫颈癌细胞的表达，而UCHL5增加XRCC6、β-catenin和TGF-βR1等分子的表达水平，调控宫颈癌细胞的糖酵解，最终增强宫颈癌细胞的辐射抗性、促进宫颈癌细胞残癌加速重建。本项目为深入阐释宫颈癌放疗抵抗提供了新的实验依据，为基于泛素化调节研发宫颈癌治疗新策略提供了理论基础。.