基本螺旋-环-螺旋家族成员 e41 (BHLHE41) 在肿瘤中经常失调，在各种癌症的恶性进展中发挥着至关重要的作用。然而，其在膀胱癌（BCa）中的具体功能和潜在机制在很大程度上仍未被探索。通过qRT-PCR和蛋白质印迹法检测BHLHE41在BCa组织和细胞中的表达水平。通过慢病毒感染构建稳定敲低或过表达BHLHE41的BCa细胞。分别采用CCK-8、流式细胞术、伤口愈合、Transwell侵袭实验检测细胞增殖、细胞周期分布、迁移和侵袭的变化。 Western blot检测相关蛋白的表达水平。通过免疫共沉淀分析探讨了 BHLHE41 和 PYCR1 之间的相互作用。在本研究中，我们发现 BHLHE41 在膀胱癌组织和细胞系中低表达，BHLHE41 的低表达与膀胱癌患者总生存期较差相关。在功能上，通过操纵BHLHE41的表达，我们通过各种体外和体内实验证明，BHLHE41的过表达显着延缓膀胱癌的细胞增殖、迁移、侵袭，并诱导细胞周期停滞，而BHLHE41的沉默则引起相反的效果。从机制上讲，我们发现BHLHE41直接与PYCR1相互作用，降低其稳定性并导致PYCR1泛素化和降解，从而失活PI3K/AKT信号通路。拯救实验表明，BHLHE41 过度表达引起的影响可以通过进一步上调 PYCR1 来减弱。BHLHE41 可能是膀胱癌中有用的预后生物标志物和肿瘤抑制因子。 BHLHE41/PYCR1/PI3K/AKT 轴可能是膀胱癌干预的潜在治疗靶点。© 2024。作者。

The basic helix-loop-helix family member e41 (BHLHE41) is frequently dysregulated in tumors and plays a crucial role in malignant progression of various cancers. Nevertheless, its specific function and underlying mechanism in bladder cancer (BCa) remain largely unexplored.The expression levels of BHLHE41 in BCa tissues and cells were examined by qRT-PCR and western blot assays. BCa cells stably knocking down or overexpressing BHLHE41 were constructed through lentivirus infection. The changes of cell proliferation, cell cycle distribution, migration, and invasion were detected by CCK-8, flow cytometry, wound healing, transwell invasion assays, respectively. The expression levels of related proteins were detected by western blot assay. The interaction between BHLHE41 and PYCR1 was explored by co-immunoprecipitation analysis.In this study, we found that BHLHE41 was lowly expressed in bladder cancer tissues and cell lines, and lower expression of BHLHE41 was associated with poor overall survival in bladder cancer patients. Functionally, by manipulating the expression of BHLHE41, we demonstrated that overexpression of BHLHE41 significantly retarded cell proliferation, migration, invasion, and induced cell cycle arrest in bladder cancer through various in vitro and in vivo experiments, while silence of BHLHE41 caused the opposite effect. Mechanistically, we showed that BHLHE41 directly interacted with PYCR1, decreased its stability and resulted in the ubiquitination and degradation of PYCR1, thus inactivating PI3K/AKT signaling pathway. Rescue experiments showed that the effects induced by BHLHE41 overexpression could be attenuated by further upregulating PYCR1.BHLHE41 might be a useful prognostic biomarker and a tumor suppressor in bladder cancer. The BHLHE41/PYCR1/PI3K/AKT axis might be a potential therapeutic target for bladder cancer intervention.© 2024. The Author(s).