Bai Yang, Zhang Zhanqiang, Bi Jiong, Tang Qian, Jiang Keying, Yao Chen, Wang Wenjian
Laboratory of Department of Surgery, the First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510080, Guangdong, China.
Department of Thyroid, the First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510080, Guangdong, China.
Cancer Cell Int. 2024 Jun 10;24(1):204. doi: 10.1186/s12935-024-03395-1.
Aberrant Derlin-1 (DERL1) expression is associated with an overactivation of p-AKT, whose involvement in breast cancer (BRCA) development has been widely speculated. However, the precise mechanism that links DERL1 expression and AKT activation is less well-studied.
Bioinformatic analyses hold a promising approach by which to detect genes' expression levels and their association with disease prognoses in patients. In the present work, a dual-luciferase assay was employed to investigate the relationship between DERL1 expression and the candidate miRNA by both in vitro and in vivo methods. Further in-depth studies involving immunoprecipitation-mass spectrum (IP-MS), co-immunoprecipitation (Co-IP), as well as Zdock prediction were performed.
Overexpression of DERL1 was detected in all phenotypes of BRCA, and its knockdown showed an inhibitory effect on BRCA cells both in vitro and in vivo. The Cancer Genome Atlas (TCGA) database reported that DERL1 overexpression was correlated with poor overall survival in BRCA cases, and so the quantification of DERL1 expression could be a potential marker for the clinical diagnosis of BRCA. On the other hand, miR-181c-5p was downregulated in BRCA, suggesting that its overexpression could be a potent therapeutic route to improve the overall survival of BRCA cases. Prior bioinformatic analyses indicated a somewhat positive correlation between DERL1 and TRAF6 as well as between TRAF6 and AKT, but not between miR-181c-5p and DERL1. In retrospect, DERL1 overexpression promoted p-AKT activation through K63 ubiquitination. DERL1 was believed to directly interact with the E3 ligase TRAF6. As Tyr77Ala or Tyr77Ala/Gln81Ala/Arg85Ala/Val158Ala attempts to prevent the interaction between DERL1 and TRAF domain of TRAF6, resulted in a significant reduction in K63-ubiquitinated p-AKT production. However, mutations in Gln81Ala, Arg85Ala, or Val158Ala could possibly interrupt with these processes.
Our data confirm that mediation of the miR-181c-5p/DERL1 pathway by TRAF6-linked AKT K63 ubiquitination holds one of the clues to set our focus on toward meeting the therapeutic goals of BRCA.
异常的Derlin-1（DERL1）表达与p-AKT的过度激活相关，其在乳腺癌（BRCA）发展中的作用已被广泛推测。然而，将DERL1表达与AKT激活联系起来的精确机制尚未得到充分研究。
生物信息学分析是一种很有前景的方法，可用于检测患者基因的表达水平及其与疾病预后的关联。在本研究中，采用双荧光素酶测定法，通过体外和体内方法研究DERL1表达与候选miRNA之间的关系。进一步进行了涉及免疫沉淀-质谱（IP-MS）、免疫共沉淀（Co-IP）以及Zdock预测的深入研究。
在BRCA的所有表型中均检测到DERL1的过表达，其敲低在体外和体内均对BRCA细胞显示出抑制作用。癌症基因组图谱（TCGA）数据库报告称，DERL1过表达与BRCA患者的总体生存率差相关，因此DERL1表达的定量可能是BRCA临床诊断的潜在标志物。另一方面，miR-181c-5p在BRCA中表达下调，这表明其过表达可能是提高BRCA患者总体生存率的有效治疗途径。先前的生物信息学分析表明DERL1与TRAF6之间以及TRAF6与AKT之间存在一定的正相关，但miR-181c-5p与DERL1之间不存在正相关。回顾来看，DERL1过表达通过K63泛素化促进p-AKT激活。DERL1被认为直接与E3连接酶TRAF6相互作用。由于Tyr77Ala或Tyr77Ala/Gln81Ala/Arg85Ala/Val158Ala试图阻止DERL1与TRAF6的TRAF结构域之间的相互作用，导致K63泛素化的p-AKT产生显著减少。然而，Gln81Ala、Arg85Ala或Val158Ala的突变可能会干扰这些过程。
我们的数据证实，通过TRAF6连接的AKT K63泛素化介导的miR-181c-5p/DERL1途径是我们聚焦于实现BRCA治疗目标的线索之一。
