Department of Urology, Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, No. 32, West Section 2, 1st Ring Road, Qingyang District, Chengdu City, 610072, Sichuan Province, China.
Sci Rep. 2024 Oct 31;14(1):26188. doi: 10.1038/s41598-024-76724-z.
Bladder cancer (BCa) is a lethal malignancy of the urinary system and exhibits a poor prognosis. Chaperonin-containing tailless complex polypeptide 1 subunit 3 (CCT3) acts as an oncogene in various tumors, whereas its effect on BCa remains unknown. We identified the ferroptosis-associated differentially expressed genes through bioinformatic analysis and selected CCT3 for further verification. The levels of cell viability, apoptosis, migration, invasion, and proliferation were measured to clarify the effect of silencing CCT3 on BCa cells. Then we evaluated the role of CCT3 knockdown in vivo. Ferroptosis was assessed by the expression detection of the ferroptosis-related proteins. The underlying mechanism was predicted by RNA sequencing and verified by an agonist for nucleotide-binding and oligomerization domain 1 (NOD1). Western blotting was conducted to detect the protein expression of NOD1, nuclear factor kappa B (NF-κB) inhibitor alpha (IκBα), and phospho-IκBα (p-IκBα). In vitro, down-regulation of CCT3 suppressed the cell viability, migration, invasion, and proliferation, as well as induced apoptosis of BCa cells. In vivo, silencing CCT3 elevated the body weight of mice and suppressed the BCa progression. In addition, CCT3 knockdown could induce ferroptosis in vitro and in vivo. CCT3 knockdown suppressed the expression of NOD1 and p-IκBα/IκBα and the NOD1 agonist could reverse the effect of CCT3 suppression on BCa in vitro and in vivo. In summary, our findings demonstrate that silencing CCT3 inhibits BCa via induction of ferroptosis and suppression of the NOD1-NF-κB pathway.
膀胱癌（BCa）是一种致命的泌尿系统恶性肿瘤，预后不良。伴侣蛋白包含的无尾复合物多肽 1 亚基 3（CCT3）在各种肿瘤中作为癌基因发挥作用，而其对 BCa 的影响尚不清楚。我们通过生物信息学分析鉴定了与铁死亡相关的差异表达基因，并选择 CCT3 进行进一步验证。通过测量细胞活力、凋亡、迁移、侵袭和增殖水平，阐明沉默 CCT3 对 BCa 细胞的影响。然后我们评估了 CCT3 敲低在体内的作用。通过铁死亡相关蛋白的表达检测评估铁死亡。通过 RNA 测序预测潜在机制，并通过核苷酸结合寡聚化结构域 1（NOD1）激动剂进行验证。通过 Western blot 检测 NOD1、核因子 kappa B（NF-κB）抑制剂 alpha（IκBα）和磷酸化 IκBα（p-IκBα）的蛋白表达。在体外，下调 CCT3 抑制了 BCa 细胞的活力、迁移、侵袭和增殖，并诱导了细胞凋亡。在体内，沉默 CCT3 提高了小鼠的体重并抑制了 BCa 的进展。此外，CCT3 敲低可在体外和体内诱导铁死亡。CCT3 敲低抑制了 NOD1 和 p-IκBα/IκBα 的表达，NOD1 激动剂可逆转 CCT3 抑制在体外和体内对 BCa 的作用。总之，我们的研究结果表明，沉默 CCT3 通过诱导铁死亡和抑制 NOD1-NF-κB 通路抑制 BCa。
